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Application of Immunohistochemistry in Thyroid Pathology.

In 2013, the American Cancer Society estimated that 60 220 new cases of thyroid carcinoma would be diagnosed in the United States, and 1850 persons would die of the disease. The overall incidence of thyroid carcinoma has increased more rapidly than that of any other malignancy in recent years, especially in women, from 1.3 per 100 000 in 1935 to 16.3 per 100 000 in 2008 (1, 2) Most thyroid carcinomas (95%) are derived from follicular epithelial cells and are mainly well differentiated, including papillary thyroid carcinoma (PTC) and follicular thyroid carcinoma (FTC), with a minor fraction of poorly differentiated carcinomas (PDCs) and undifferentiated carcinomas (UDCs). (3) Only 3% of thyroid carcinomas are medullary carcinomas (MCs) derived from parafollicular C cells.

The diagnosis of thyroid carcinomas is straightforward in most cases. However, pathologists are not infrequently confronted with lesions exhibiting equivocal features that make the distinction of benign from malignant difficult. Although the diagnostic criteria for PTC and FTC are clearly outlined in textbooks, the application of the criteria is subjective and circumstantial. Interobservor or intra-observor disagreements in the diagnosis of follicular thyroid lesions are well documented, even among expert pathologists. (4-7) In recent years, tremendous efforts in search of objective measures that differentiate benign from malignant lesions have been undertaken, mainly involving immunohisto chemical markers and molecular markers, using somatic mutation, gene expression, and microRNA analyses. The immunohistochemical markers are the scope of this review. The objective of this article is to assess the diagnostic utility of the most commonly studied and published immunomarkers in the current literature in the field of thyroid pathology.

IMMUNOMARKERS USED IN THYROID-ORGAN SPECIFIC DIFFERENTIATION

Thyroid transcription factors (TTFs) TTF1, paired box gene 8 (PAX8), and TTF2 (FOXE1) are crucial for thyroid organogenesis and differentiation. (8-14) The transcription factors control the expression of thyroglobulin (TGB), thyroperoxidase (TPO), thyroid-stimulating hormone receptor, and thyroid iodine transporter. (15, 16) Immunohisto-chemically, they serve as organ-specific immunomarkers. Thyroglobulin is the primary synthetic product of the thyroid and the macromolecular precursor of thyroid hormones (T3 and T4), serving as a specific marker for thyroid follicular cell origin. (17-19)

Thyroid Transcription Factor 1

Thyroid transcription factor 1, also named NKX2 homeobox 1 (NKX2.1), is a nuclear protein, approximately 38 kDa, composed of a single polypeptide of 371 amino acids belonging to the family of homeodomain transcription factors. Thyroid transcription factor 1 plays a crucial role in the organogenesis and differentiation of thyroid and lung. (9-11) In addition, TTF1 transcripts were detected in developing diencephalon, restricted to the hypothalamic area and to the infundibulum at the earliest stages of their differentiation, forming the posterior lobe of the pituitary, namely, the neurohypophysis. (9, 20)

Thyroid transcription factor 1 expression by immunohisto-chemical analysis was initially exclusively identified in thyroid and lung epithelial tissues, including normal, benign, and malignant tissues. (18, 21-30) In routine practice, TTF1 became one of the most commonly used immunomarkers to identify thyroid or lung primary tumor in the setting of metastasis and to differentiate adenocarcinoma from squamous cell carcinoma in poorly differentiated non-small cell carcinomas of the lung in small biopsy or cytologic specimens. In recent years, TTF1-positive, non-pulmonary, nonthyroid carcinomas have been reported, including colorectal, ovarian, breast, endometrial, and endocervical adenocarcinomas. (31-42) Comparing the 2 commercially available TTF1 monoclonal antibodies, SPT24 and 8G7G3/1, the rate of aberrant TTF1 expression in nonpulmonary, nonthyroid carcinomas is higher for SPT24, greater than 10% in colorectal adenocarcinomas as reported by some investigators. However, if using the 8G7G3/1 clone, the aberrant expression rate is low, less than 2% in general.

Normal thyroid follicular cells and parafollicular cells show diffuse expression of TTF1. In thyroid neoplasm, TTF1 expression was reported in nearly 100% of PTCs, FTCs, and follicular adenomas (FAs); in approximately 90% of PDCs and MCs; and in none to fewer than 25% of UDCs. *

Paired Box Gene 8

Paired box gene 8 is a member of the paired box (PAX) family of transcription factors that is expressed during organogenesis of the thyroid gland, mullerian tract, and kidney. (47-52) Expression of PAX8 was mainly reported in thyroid and renal neoplasms and infrequently in bladder tumors, as well as mullerian-origin malignancies (including ovarian malignant neoplasms). (26, 46, 53-65) Few studies (53, 54) also reported its expression in thymic and parathyroid neoplasms. Recently, PAX8 expression was documented in pancreatic neuroendocrine tumors. (66-68) Tacha et al (54) reported that other cancers, including carcinomas of breast, lung, prostate, gastrointestinal tract, liver, mesothelioma, melanoma, and testicular tumors, lacked PAX8 expression; 14% (3 of 22) of pancreatic adenocarcinomas and 40% (2 of 5) of rhabdomyosarcomas were PAX8 positive.

In normal tissues, strong nuclear staining for PAX8 was observed in follicular cells of the thyroid; mullerian epithelial cells including endometrium, endocervix, and secretory cells of the fallopian tube; renal tubular epithelium; epithelial lining of the vas deferens; as well as islet cells of the pancreas. (54) Diffuse weak to moderate nuclear staining was observed in some cases of parathyroid tissue and nonneo-plastic thymic epithelial cells. Urothelium and squamous mucosa were reported to have weak and patchy staining. Several studies investigated PAX8 expression in thyroid neoplasms, reporting a positive rate of nearly 100% in PTCs and FAs, (43, 46, 53, 54, 69) 91% to 100% in FTCs, (46, 53, 54) 75% to 100% in PDCs, (46, 53, 58) and 50% to 80% in UDCs. (46, 53, 54, 56, 58)

Medullary carcinomas were reported to be PAX8 nonreactive with rare exceptions: positive in 75% (6 of 8, most 1+) (46) and 41% (13 of 32) (54) of cases.

Thyroid Transcription Factor 2

Thyroid transcription factor 2, also named forkhead box E1 (FOXE1), was originally identified as a thyroid-specific, forkhead-domain-containing nuclear protein capable of recognizing and binding to a DNA sequence present in the promoters of both TGB and TPO, 2 genes expressed exclusively in thyroid follicular cells. (15, 70) Thyroid transcription factor 2 expression is restricted to the thyroid; anterior pituitary; epithelia of the oropharynx, trachea, and esophagus; exocrine cells of the seminiferous tubules of the testis; and epidermis and hair follicles. (12, 71-78) Thyroid transcription factor 2 is 1 of 3 TTFs that play critical roles in thyroid organogenesis and differentiation. However, TTF2 seems to play a role in the development of the negative controller of thyroid-specific gene expression. (79, 80) Mutations in TTF2 (FOXE1) may be involved in human disorders characterized by congenital hypothyroidism, thyroid dysgenesis, and cleft palate. (81, 82)

There are only few studies in literature investigating TTF2 expression by immunohistochemical analysis in human tumors and normal tissues. Nonaka et al (46) reported that TTF2 was expressed exclusively in normal thyroid follicular cells and a few C cells in thyroid C-cell hyperplasia and thyroid neoplasms, including 100% of cases of PTC (17 of 17), FA (18 of 18), FTC (16 of 16), and PDC (7 of 7); 75% (6 of 8) of cases of MC in a focal staining pattern; and only 7% (2 of 28, focal 1+, 1%-25% of nuclei) of cases of UDC. All other neoplastic and nonneoplastic tissues showed no reactivity, including lung, esophagus, stomach, colon, pancreas, kidney, breast, ovary, prostate, urinary bladder, skin, testis, lymph node, and soft tissue. Matoso et al (83) studied 6 cases of spindle cell foci in thyroid glands by immunohistochemical analysis and found that 100% (6 of 6) of cases showed nuclear staining for TTF2. Increased expression of TTF2 by in situ hybridization was reported in epidermis and basal cell carcinomas. Zhang et al (36) documented the lack of expression of TTF2 by immunohisto-chemical evaluation in 212 cases of ovarian and endometrial malignancies to ascertain that the TTF1-positive tumors in their study were indeed not follicular cell in origin.

Although there are only few studies evaluating TTF2 expression in neoplastic and nonneoplastic tissues by immunohistochemistry, the data are appealing. Further study in a large series of cases is deemed necessary to assess the diagnostic utility of TTF2 in routine practice.

Thyroglobulin

Thyroglobulin, a thyroid hormone precursor, is a glycoprotein synthesized by thyrocytes, transported to the apical surface and secreted into the follicles, constituting the major component of colloid. Immunohistochemically, TGB expression is diffuse in 100% of normal thyroid follicular epithelial cells and 83% to 100% of FAs. (18, 43, 84) However, the staining pattern in lesions with Hurthle cell morphology was reported as weak or focal. In primary thyroid carcinomas, the expression of TGB, both at the messenger RNA and protein level, showed a certain degree of correlation with tumor differentiation. Studies (18, 43, 84-88) have reported 100% expression of TGB in PTC, 75% to 96% in FTC, 57% to 92% in PDC, and near lack of expression in UDC and MC. Metastatic thyroid carcinomas were reported as showing staining patterns similar to those of the primary tumors. (18, 89) In nonthyroid tissues and tumors, including parathyroid gland, lung, stomach, pancreas, ovary, kidney, colon, salivary glands, prostate, breast, and glomus body, TGB expression was lacking. (43, 89-91) In our previous study, (92) TGB expression was demonstrated in 14 of 14 (100%) cases of normal thyroid tissue, 45 of 45 (100%) cases of PTC, 36 of 36 (100%) cases of FTC, and 0 of 10 (0%) cases of MC.

IMMUNOMARKERS USED IN THE DIFFERENTIAL DIAGNOSIS OF THYROID NEOPLASMS

The histomorphologic diagnosis of thyroid neoplasm remains the cornerstone in the classification of thyroid follicular lesions. However, for those tumors that are poorly differentiated or undifferentiated, not follicular derived, and exhibit equivocal histomorphologic features, the application of immunohistochemical biomarkers may play an active or complementary role in their accurate classification. (93-99) Among the variety of biomarkers reported in the literature, Hector Battifora mesothelial-1 (HBME-1), galectin-3 (GAL3), cytokeratin 19 (CK19), Cbp/p300-interacting transactivator with Glu/Asp-rich carboxy-terminal domain, 1 (CITED1), and TPO are most promising. However, there is no single marker sensitive enough to provide a definitive malignant diagnosis. Therefore, different panels of combined immunomarkers were proposed by many investigators. (98, 100-112) The combination of HBME-1, GAL-3, and CK19 was by far the most common panel evaluated by investigators, and their diffuse expression has not been reported in benign lesions. Other markers, such as antibodies against proteins involved in cell proliferation and in the regulation of cell cycle, as well as oncogene proteins, are also being studied and documented in the literature. Recently, we found a distinct membranous staining pattern for trophoblastic cell surface antigen 2 (TROP2), a 35-kDa type 1 transmembranous glycoprotein, in PTCs; in contrast, follicular neoplasms (FAs and FTCs) were nonreactive or showed only rare focal, weak cytoplasmic staining. We propose that TROP2 is a potential novel immunomarker for the identification of PTC that can be used in a panel to increase diagnostic accuracy when encountering a difficult follicular cell-derived lesion. (113)

Hector Battifora Mesothelial-1

Hector Battifora mesothelial-1 is an unelucidated membrane antigen found in the microvilli of mesothelial cells, normal tracheal epithelium, and adenocarcinoma of the lung, pancreas, and breast. In the past decade, several studies have investigated HBME-1 expression in benign and malignant thyroid tissues; the published studies are reviewed and summarized in Table 1. In normal thyroid tissue, there was virtually no expression of HBME (104, 108, 114-116) Overexpression of HBME-1 was demonstrated in malignant thyroid neoplasms, especially PTCs, with the exception of Hurthle cell carcinomas. Several investiga tors (104,108,114-119) have reported a reduced or lack of expression for HBME-1 in Hurthle cell carcinomas or thyroid neoplasms with Hurthle cell features. Studies suggested that overexpression of HBME-1 in a thyroid nodule was an indicator of malignancy, especially true for PTC. The overall sensitivity of HBME-1 was 78.8% for thyroid malignancy, 87.3% for PTC, and 65.2% for FTC. The specificity was 82.1%.+ Expression of HBME-1 was also noted in benign thyroid lesions such as FA, nodular goiter, and lymphocytic thyroiditis (LTis), usually in a focal staining fashion, with a reported overall positive rate of 26%, 12%, and 19%, respectively. ([dagger]) Hyaline trabecular tumors were reported to lack HBME-1 expression, suggesting its discriminating role in the distinction between hyaline trabecular tumor and PTC; however, Lenggenhager et al (119) recently reported that 37.5% (3 of 8) of cases of hyaline trabecular tumor showed patchy but strong membranous and/or cytoplasmic reactivity to HBME-1. Many investigators proposed using a panel of immunomarkers, most commonly a combination of HBME-1, CK19, and GAL-3, to increase the discriminating power between benign and malignant neoplasms when a histologically equivocal lesion is encountered.

Cytokeratin 19

Cytokeratin 19 is a low-molecular-weight cytokeratin found in a variety of simple or glandular epithelia, both normal and their neoplastic counterparts. In the thyroid gland, normal follicular epithelium usually has shown no detectable CK19 expression (110, 115, 129); however, few re ports (114, 128, 130) noted CK19 expression in normal thyroid tissue in a focal staining pattern, especially in inflamed tissue. Many studies reported a strong and diffuse staining pattern of CK19 in PTC. However, its expression in follicular cells of LTis (114, 115) and follicular neoplasms (FA or FTC) was also demonstrated8; therefore, positive CK19 staining lacks specificity for PTC or malignancy. Table 2 summarizes the data from studies of CK19 expression in various thyroid lesions. The overall sensitivity of CK19 was 79.3% for malignancy, 82.2% for PTC, and 44.3% for FTC. The specificity was 63.1%. Overexpression of CK19 is a good indicator for PTC; however, the sensitivity for follicular carcinoma is low. There are high rates of reactivity in benign thyroid lesions, especially in LTis. (114, 115) Compared with the diffuse, strong reactivity in PTC, most studies indicated focal reactivity in benign lesions. Cytokeratin 19 may have added value as part of a panel of immunomarkers in the diagnosis of PTC.

Galectin-3

Galectin-3 is a member of a family of [beta]-galactoside-binding animal lectins shown to be involved in tumor progression and metastasis. Both nuclear and cytoplasmic expression of GAL-3 has been demonstrated in a variety of tissues and cells. Study of mouse 3T3 fibroblasts revealed the presence of both phosphorylated and nonphosphorylated GAL-3; the former resides in both nucleus and cytoplasm; the latter, exclusively in the nucleus. Cell proliferation is associated with an increased level of both forms, while alterations in nuclear versus cytoplasmic GAL-3 localization have been shown to be associated with neoplastic progression. (136, 137) Galectin-3 plays an important role in cell-cell/cell-matrix adhesion, cell growth, neo-plastic transformation/spread, cell cycle regulation/apoptosis, and cell repair processes. In recent years, overexpression of GAL-3 has been reported in various human carcinomas, most noticeably in well-differentiated follicular-derived thyroid carcinomas. Many studies ([parallel]) investigated GAL-3 expression in cytologic and histologic specimens of thyroid nodular lesions; these are summarized in Table 3. Galectin-3 has been found to be useful in differentiating malignant thyroid lesions (such as PTC and the follicular variant of PTC) from benign lesions. The overall sensitivity of GAL-3 was 84.6% for malignancy, 87.5% for PTC, and 72.6% for FTC. The specificity was 83.6%. In general, GAL-3 expression in benign lesions is often focal, and in contrast, is diffuse in malignant lesions.

Trophoblastic Cell Surface Antigen 2

Trophoblastic cell surface antigen 2, also known as tumor-associated calcium signal transducer 2 (TAC-STD2), is a transmembrane glycoprotein associated with tumor development and progression in a variety of epithelial carcinomas including ovarian, colorectal, pancreatic, gastric, pulmonary, endometrioid endometrial, and oral cavity squamous cell carcinomas. (147-157) In normal tissues, TROP2 expression was reported as low or lacking. Many studies (158-163) have demonstrated that TROP2 overexpression is associated with tumor aggressiveness and poor prognosis.

Immunohistochemical evaluation of TROP2 expression in thyroid tumors has not been well studied. Recently, we (113) evaluated TROP2 expression in 136 cases of thyroid neoplasms (48 cases of PTC, 37 cases of FTC, 51 cases of FA) and normal thyroid tissues (n = 15) on tissue microarray (TMA) sections, as well as in routine sections of 61 atypical follicular lesions and 20 benign thyroid lesions (10 cases each of chronic LTis and nodular goiter) by immunohisto-chemical analysis with the Ventana BenchMark Ultra (Ventana Medical Systems Inc, Tucson, Arizona). In the TMAs of thyroid neoplasms, 90% (43 of 48) of PTCs demonstrated a strong membranous staining pattern, with most being diffuse (3+ or 4+), as illustrated in Figure 1, A and B. In contrast, 96% (49 of 51) of FAs and 89% (33 of 37) of FTCs lacked TROP2 expression; only 2 of 51 FA cases and 4 of 37 FTC cases showed focal (1+) strong cytoplasmic staining without membranous condensation, as illustrated in Figure 1, C and D. The staining results are summarized in Table 4. For the 61 routine sections of atypical follicular lesion, 70% (23 of 33) of PTCs were positive for TROP2, with a diffuse staining pattern (3+ or 4+) in 83% (19 of 23) of the positive cases. All 11 cases of atypical follicular neoplasm and 17 cases of adenomatoid nodule with focal nuclear atypia showed lack of TROP2 expression. The staining results are summarized in Table 5. The normal thyroid tissue on TMA revealed weak cytoplasmic staining; no membranous staining pattern was identified. The 10 surgical cases each of nodular hyperplasia and chronic LTis showed no TROP2 expression, except rare weak to moderate membranous staining in the lining cells of a degenerative cyst in 1 of the 10 cases of chronic LTis, as illustrated in Figure 1, E and F. Our data suggest that TROP2 is a potential novel immunomarker for identification of both classic and follicular variants of PTC. Based on our study, TROP2 appears to be a more specific marker than the 3 traditional markers (CK19, HBME-1, and GAL-3); however, additional studies in a large number of difficult cases from multiple institutions are warranted to substantiate the current findings.

Thyroperoxidase

Thyroperoxidase is a thyroid-specific enzyme reflecting normal thyroid function. Thyroperoxidase expression is demonstrated in normal thyroid follicular epithelial cells, usually in diffuse fashion. (103, 164) During thyroid cell dedifferentiation, the expression of TPO is lost. Therefore, lack of TPO expression is regarded as a marker of malignancy.

Several studies (103, 120, 164-169) investigated the diagnostic utility of TPO in thyroid lesions, yielding a sensitivity of 90% for PTC and 76% for FTC and a specificity of 88%. Table 6 summarizes the data from reported studies of TPO expression in thyroid lesions. The earlier studies produced more promising results than recent ones. Thyroperoxidase appears highly sensitive for PTC but borderline to poorly sensitive for FTC. The staining patterns of FTC and FA showed a certain degree of overlapping without meaningful discriminatory value. A couple of recent studies (103, 129) used a combination of TPO and GAL-3 to improve diagnostic accuracy. They yielded a sensitivity of 96% to 98% for PTC and 44% to 67% for FTC, proposing that the application of TPO immunostain in combination with other biomarkers is useful, and with added value in the diagnosis of PTC.

OTHER CELL ADHESION MOLECULES, CELL CYCLE REGULATORY PROTEINS, AND ONCOGENES

E-cadherin, a calcium-dependent transmembrane cell adhesion molecule, is required for normal epithelial function. Down-regulation of E-cadherin expression has been observed in various carcinomas and is usually associated with an advanced stage and progression. In normal and benign thyroid lesions, high expression of E-cadherin was demonstrated and restricted to thyrocytes. (170-172) In thyroid carcinomas, E-cadherin expression was reduced in PDC, lost in UDC, and preserved in most minimally invasive carcinomas. (170-175) These studies suggest that loss of E-cadherin expression is the crucial event in dedifferentiation, progression, and metastatic spread of thyroid carcinomas.

Fibronectins (FNs), extracellular matrix proteins produced by fibroblasts, are involved in cell adhesion, migration, and tumor progression. Oncofetal FNs, isoforms of FN, are highly expressed in fetal and neoplastic tissues, especially PTCs. (176-180) Prasad et al (108) investigated the expression of

FN-1, CITED-1, HBME-1, and CK19 in various thyroid tissues by immunohistochemical analysis and revealed the expression of FN-1 in 91% (61 of 67) of PTCs, 100% (4 of 4) of UDCs, 50% (3 of 6) of FTCs, and 75% (6 of 8) of Hurthle cell carcinomas; in contrast, none of the normal (n = 59) or benign thyroid lesions (n = 43) expressed FN-1, with the exception of 2 of 29 cases of nodular goiter, which showed cytoplasmic and membranous expression of FN-1 in thyrocytes in association with fresh hemorrhage and fibrin deposition. Liu et al (109) evaluated a panel of immuno-markers on 100 benign thyroid tissues and 77 thyroid carcinomas on TMAs, revealing overexpression of FN-1 in 96% of PTCs and 86% of FTCs, suggesting that a panel consisting of GAL-3, FN-1, and intracellular sodium/iodide symporter demonstrates 98% accuracy in differentiating FA from malignant thyroid follicular lesions.

Cluster of differentiation (CD) 44v6 is the isoform of CD44, a cell surface membrane glycoprotein that plays a role in the regulation of cell-cell and cell-matrix interactions as well as cell migration. (181) Expression of CD44v6 has been reported in several carcinomas. (182-185) Few studies investigating CD44v6 expression in thyroid lesions found overexpression of CD44v6 in well-differentiated PTCs and FTCs, suggesting the potential utility of CD44v6 in combination with GAL-3 in distinguishing benign from malignant thyroid neoplasms. (186-191) However, CD44v6 expression was also demonstrated in benign lesions and poorly differentiated or undifferentiated thyroid carcinomas at a lower rate of expression.

Cyclin-dependent kinase inhibitor 1B (p27Kip1), a nuclear cyclin-dependent kinase inhibitor, plays a major role in controlling progression from the G1 to the S phase of the cell cycle. The loss of regulatory control of the cell cycle, leading to unrestrained cell proliferation, is the hallmark of cancers. Down-regulation of p27Kip1 has been shown to correlate with histologic loss of differentiation or high-grade morphology in various human carcinomas (192-196) and is emerging as an important prognostic factor. Studies (197, 198) revealed strong nuclear reactivity in normal thyrocytes. In thyroid neoplasms, p27Kip1 expression was significantly higher in benign lesions than malignant thyroid neoplasms, suggesting the potential utility of p27Kip1 in the differentiation of benign from malignant follicular lesions. (197-201)

Cyclin D1, a member of the family of cyclins, is a 36-kDa nuclear protein that functions as the regulatory subunit of cyclin-dependent kinases (CDK4 and CDK6) and controls the progression of the G1/S phases of the cell cycle. (202) Deregulation of cyclin expression results in the loss of control of normal cell growth and oncogenesis. Over-expression of cyclin D1 has been demonstrated in various human cancers, including thyroid carcinomas. (201,203-209)

Normal thyrocytes are immunohistochemically negative for cyclin D1. Studies have been conducted to investigate the expression of cyclin D1 in thyroid neoplasms. Melck et al (210) evaluated the expression of cell cycle regulators, including cyclin D1, in TMAs of 100 benign and 105 malignant thyroid lesions. They demonstrated over-expression of cyclin D1 in 87.1% of malignant and 45.7% of benign thyroid lesions; similar findings were reported by Seybt et al (211) and Park et al. (102) Wang et al (212) investigated cyclin D1 expression in 34 conventional PTCs, 10 minimally invasive FTCs, and 32 more aggressive thyroid carcinomas. Their study demonstrated overexpression of cyclin D1 in most aggressive thyroid carcinomas. Erickson et al (213) evaluated the expression of Ki-67 and cyclin D1 in Hurthle cell neoplasms (59 Hurthle cell adenomas, 55 Hurthle cell carcinomas, and 14 Hurthle cell neoplasms of uncertain malignant potential). This revealed overexpression of cyclin D1 in 18% of Hurthle cell carcinomas, compared with only 1.7% of the adenomas and none of the uncertain malignant potential tumors. It has been well demonstrated that cyclin D1 is overexpressed in malignant thyroid lesions compared to benign lesions; however, the staining patterns showed heterogeneity, which limits its utility as a diagnostic marker.

[beta]-catenin, a 92-kDa multifunctional protein, plays an important role in cell adhesion and signal transduction and serves as a downstream effector in the Wnt signaling pathway. (214) In normal resting cells, [beta]-catenin forms cytoplasmic/membranous-bound complexes with E-cadherin. Upon activation, [beta]-catenin translocates to the nucleus, promoting tumor growth through activation of the Wnt signaling pathway. (93,215-217) Normal thyroid follicular cells display a strong membranous immunoreactivity for bcatenin. (218, 219) Studies were conducted to investigate the expression and the pattern of expression of b-catenin in various thyroid tumors. Garcia-Rostan et al (218) reported nuclear expression of b-catenin in 32.1% (9 of 28) of PDCs, 79.3% (23 of 29) of UDCs, and 0% (0 of 58) of the well-differentiated carcinomas. Rossi et al (120) reported [beta]-catenin expression in 80% (12 of 15) of PDCs but 0% (0 of 9) of UDCs. Ishigaki et al (220) observed cytoplasmic immuno-reactivity for [beta]-catenin in 67% (52 of 78) of PTCs, 25% (5 of 20) of FTCs, and only 9% (3 of 34) of FAs. The association of aberrant nuclear [beta]-catenin expression and poor prognosis was observed by some investigators. (218)

p53 is a tumor suppressor gene product that plays an important role in normal cell growth. Mutations of the TP53 gene lead to accumulation of p53, which can be detected by immunohistochemistry. p53 expression has been reported in various tumors, mainly in UDCs and PDCs and rarely in well-differentiated carcinomas as well as MCs in thyroid neoplasms. (220-226)

BRAF Mutation-Specific Antibody

The BRAF (the B-isoform of RAF kinase) oncogene is mutated in several types of tumors, such as colorectal adenocarcinoma, PTC, glioma, gastrointestinal adenocarcinoma, melanoma, and pulmonary adenocarcinoma. (227) The most common mutation in BRAF is due to a T to A switch at position 1796, which results in an alteration from valine to glutamate at the V600E. The BRAF V600E point mutation has been reported in approximately 50% of PTCs, with a higher frequency (approximately 70%) in tall cell variant and oncocytic variant of PTC and a much lower frequency (approximately 20%) in follicular variant of PTC. (228) The BRAF V600E mutation is generally negative in benign follicular lesions, normal thyroid tissue, medullary carcinoma, and follicular carcinoma. A recent meta-analysis of 5655 patients suggested PTC with the BRAF mutation is associated with a higher risk of recurrent, persistent disease, lymph node metastasis, and extrathyroidal extension. (229) Many molecular techniques have been used to detect the BRAF V600E point mutation, including single-strand conformation polymorphism, mutation-specific polymerase chain reaction, direct gene sequencing, and colorimetric mutation analysis. These methods tend to be expensive, time-consuming, labor-intensive, and difficult to validate and implement in some clinical settings.

Recently, 2 mutation-specific antibodies against BRAF V600E have become commercially available; one is VE1 clone (Spring Bioscience, Pleasanton, California) and the other is anti-B-Raf mouse monoclonal antibody (New East Bioscience, Malvern, Pennsylvania). Most studies used VE1 clone, and only rare studies used anti-B-Raf mouse monoclonal antibody. (227, 228, 230-233) In general, BRAF mutation-specific antibody has been shown to be useful in detection of the BRAF V600 mutation, with a sensitivity and specificity of more than 95% when compared to other molecular methods. (227, 228, 230-233) In fact, some studies (228, 231) suggested that anti-BRAF mutation-specific antibody may be more sensitive than molecular testing in detecting the BRAF mutation. Data from selected studies of BRAF mutation-specific antibodies to detect the BRAF V600E mutation in PTCs, with the correlating molecular testing results, are summarized in Table 7. An example of the BRAF mutation in a papillary thyroid microcarcinoma, detected by immunohistochemistry using the VE1 clone, is shown in Figure 2, A through D.

CONCLUSIONS

Cytomorphology or histomorphology remains the cornerstone for the diagnosis of thyroid lesions. However, not infrequently, we encountered cases with equivocal morphology that posed a great challenge in reaching an accurate diagnosis. Ancillary studies, including somatic mutation testing, messenger RNA gene expression platforms, protein immunohistochemistry, and microRNA panels, are becoming increasingly important. Immunohistochemistry by far is the most commonly used method to complement morphologic assessment. By reviewing the published data in the current literature, we have determined that there is no individual biomarker having sufficient sensitivity or specificity to distinguish benign from malignant lesions. However, HBME-1 is often strongly and diffusely expressed in PTC; it can be used as a marker (in a panel) to aid in the PTC diagnosis. Galectin-3 (in a panel) can be useful in differentiating malignant from benign thyroid lesions; CK19 has low sensitivity and specificity. A panel of immunomarkers, as proposed by many investigators, is recommended when working on challenging thyroid follicular-derived lesions to improve diagnostic accuracy. The most commonly proposed panel is GAL-3, HBME-1, and CK19. We also recommend including TROP2 in the diagnostic panel. Other panels, such as the combination of 2 of the 3 markers or the addition of FN-1 or CITED-1, are also proposed. The application of a panel of immunomarkers improves the differential power of individual markers and aids in the accurate classification of challenging thyroid follicular-derived lesions.

The authors would like to thank Melissa Erb, AAS, for her outstanding secretarial support, Tina Brosious, HT(ASCP), and Erin Powell, HT(ASCP), for construction of TMA blocks and cutting TMA sections, Jianhui Shi, MD, PhD, and Angie Bitting, HT(ASCP), QIHC, for their assistance with immunostains, and Kathy Fenstermacher, BA, for editing this manuscript.

Please Note: Illustration(s) are not available due to copyright restrictions.

References

(1.) National Cancer Institute. SEER Stat Fact Sheets: thyroid cancer. http://seer. cancer.gov/statfacts/html/thyro.html. Accessed May 21, 2013.

(2.) Morris LG, Sikora AG, Tosteson TD, Davies L. The increasing incidence of thyroid cancer: the influence of access to care. Thyroid. 2013; 23(7):885-891.

(3.) DeLellis RA, Lloyd RV, Heitz PU, et al, eds. Pathology and Genetics of Tumours of Endocrine Organs. Lyon, France: IARC Press; 2004. World Health Organization Classification of Tumours; vol 8.

(4.) Hirokawa M, Carney JA, Goellner JR, et al. Observer variation of encapsulated follicular lesions of the thyroid gland. Am J Surg Pathol. 2002; 26(11):1508-1514.

(5.) Franc B, de la Salmoniere P, Lange F, et al. Interobserver and intra-observer reproducibility in the histopathology of follicular thyroid carcinoma. Hum Pathol. 2003; 34(11):1092-1100.

(6.) Lloyd RV, Erickson LA, Casey MB, et al. Observer variation in the diagnosis of follicular variant of papillary thyroid carcinoma. Am J Surg Pathol. 2004; 28(10):1336-1340.

(7.) Elsheikh TM, Asa SL, Chan JK, et al. Interobserver and intraobserver variation among experts in the diagnosis of thyroid follicular lesions with borderline nuclear features of papillary carcinoma. Am J Clin Pathol. 2008; 130(5):736-744.

(8.) Mitchell PJ, Tijan R. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science. 1989; 245(4916):371-378.

(9.) Kimura S, Hara Y, Pineau T, et al. The T/ebp null mouse: thyroid-specific enhancer-binding protein is essential for the organogenesis of the thyroid, lung, ventral forebrain, and pituitary. Genes Dev. 1996; 10(1):60-69.

(10.) Gehring WJ. Homeo boxes in the study of development. Science. 1987; 236(4806):1245-1252.

(11.) Guazzi S, Price M, De Felice M, Damante G, Mattei MG, Di Lauro R. Thyroid nuclear factor 1 (TTF-1) contains a homeodomain and displays a novel DNA binding specificity. EMBO J. 1990; 9(11):3631-3639.

(12.) Civitareale D, Lonigro R, Sinclair AJ, et al. A thyroid-specific nuclear protein essential for tissue-specific expression of the thyroglobulin promoter. EMBO J. 1989; 8(9):2537-2542.

(13.) Endo T, Kaneshige M, Nakazato M, et al. Thyroid transcription factor-1 activates the promoter activity of rat thyroid Na/I- symporter gene. Mol Endocrinol. 1997; 11(11):1747-1755.

(14.) Zannini M, Francis-Lang H, Plachov D, Di Lauro R. Pax 8, a paired domain-containing protein, binds to a sequence overlapping the recognition site of a homeodomain and activates transcription from two thyroid-specific promoters. Mol Cell Biol. 1992; 12(9):4230-4241.

(15.) Damante G, Di Lauro R. Thyroid-specific gene expression. Biochim Biophys Acta. 1994; 1218(3):255-266.

(16.) Dai G, Levy O, Carrasco N. Cloning and characterization of the thyroid iodine transporter. Nature. 1996; 379(6654):458-460.

(17.) Suzuki K, Mori A, Lavaroni S, et al. Thyroglobulin regulates follicular function and heterogeneity by suppressing thyroid-specific gene expression. Biochimie. 1999; 81(4):329-340.

(18.) Bejarano PA, Nikiforov YE, Swenson ES, Biddinger PW. Thyroid transcription factor-1, thyroglobulin, cytokeratin 7, and cytokeratin 20 in thyroid neoplasms. Appl immunohistochem Mol Morphol. 2000; 8(3):189-194.

(19.) Ratnatnga N, Ramadasa S. Immunohistochemical staining for thyroglobulin in poorly differentiated carcinoma of the thyroid. Ceylon Med J. 1993; 38(3): 113-116.

(20.) Lazzaro D, Price M, de Felice M, Di Lauro R. The transcription factor TTF1 is expressed at the onset of thyroid and lung morphogenesis and in restricted regions of the foetal brain. Development. 1991; 113(4):1093-1104.

(21.) Whitsett JA, Glasser SW. Regulation of surfactant protein gene transcription. Biochim Biophys Acta. 1998; 1408(2):303-311.

(22.) Ghaffari M, Zeng X, Whitsett JA, Yan C. Nuclear localization domain of thyroid transcription factor-1 in respiratory epithelial cells. Biochem J. 1997; 328(pt 3):757-761.

(23.) Boggaram V. Thyroid transcription factor-1 (TTF-1/Nkx2.1/TITF1) gene regulation in the lung. Clin Sci (Lond). 2009; 116(1):27-35.

(24.) Kondo T, Nakazawa T, Ma D, et al. Epigenetic silencing of TTF-1/NKX2-1 through DNA hypermethylation and histone H3 modulation in thyroid carcinomas. Lab invest. 2009; 89(7):791-799.

(25.) Holzinger A, Dingle S, Bejarano PA, et al. Monoclonal antibody to thyroid transcription factor-1: production, characterization, and usefulness in tumor diagnosis. Hybridoma. 1996; 15(1):49-53.

(26.) Fabbro D, Di Loreto C, Beltrami CA, Belfiore A, Di Lauro R, Damante G. Expression of thyroid-specific transcription factors TTF-1 and PAX-8 in human thyroid neoplasms. Cancer Res. 1994; 54(17):4744-4749.

(27.) Joba W, Spitzweg C, Schriever K, Heufelder AE. Analysis of human sodium/iodide symporter, thyroid transcription factor-1, and paired-box-protein-8 gene expression in benign thyroid diseases. Thyroid. 1999; 9(5):455-466.

(28.) Katoh R, Kawaoi A, Miyagi E, et al. Thyroid transcription factor-1 in normal, hyperplastic, and neoplastic follicular thyroid cells examined by immunohistochemistry and nonradioactive in situ hybridization. Mod Pathol. 2000; 13(5):5 70-576.

(29.) Ordonez NG. Thyroid transcription factor-1 is a marker of lung and thyroid carcinomas. Adv Anat Pathol. 2000; 7(2):123-127.

(30.) Kaufmann O, Dietel M. Thyroid transcription factor-1 is the superior immunohistochemical marker for pulmonary adenocarcinomas and large cell carcinomas compared to surfactant proteins A and B. Histopathology. 2000; 36(1):8-16.

(31.) Ordonez NG. Value of thyroid transcription factor-1 immunostaining in tumor diagnosis: a review and update. Appl immunohistochem Mol Morphol. 2012; 20(5):429-444.

(32.) Sullivan PS, Maresh EL, Seligson DB, et al. Expression of thyroid transcription factor-1 in normal endometrium is associated with risk of endometrial cancer development. Mod Pathol. 2012; 25(8):1140-1148.

(33.) Klingen TA, Chen Y, Suhrke P, Stefansson IM, Gundersen MD, Akslen LA. Expression of thyroid transcription factor-1 is associated with a basal-like phenotype in breast carcinomas. Diagn Pathol. 2013; 8:80.

(34.) Sakurai A, Sakai Y, Yatabe Y. Thyroid transcription factor-1 expression in rare cases of mammary ductal carcinoma. Histopathology. 2011; 59(1):145-148.

(35.) Niu HL, Pasha TL, Pawel BR, LiVolsi VA, Zhang PJ. Thyroid transcription factor-1 expression in normal gynecologic tissues and its potential significance. int J Gynecol Pathol. 2009; 28(4):301-307.

(36.) Zhang PJ, Gao HG, Pasha TL, Litzky L, Livolsi VA. TTF-1 expression in ovarian and uterine epithelial neoplasia and its potential significance, an immunohistochemical assessment with multiple monoclonal antibodies and different secondary detection systems. int J Gynecol Pathol. 2009; 28(1):10-18.

(37.) Kubba LA, McCluggage WG, Liu J, e tal. Thyroid transcription factor-1 expression in ovarian epithelial neoplasms. Mod Pathol. 2008; 21(4):485-490.

(38.) Siami K, McCluggage WG, Ordonez NG, et al. Thyroid transcription factor-1 expression in endometrial and endocervical adenocarcinomas. Am J Surg Pathol. 2007; 31(11):1759-1763.

(39.) Ye J, Findeis-Hosey JJ, Yang Q, et al. Combination of napsin A and TTF-1 immunohistochemistry helps in differentiating primary lung adenocarcinoma from metastatic carcinoma in the lung. Appl immunohistochem Mol Morphol. 2011; 19(4):313-317.

(40.) Comperat E, Zhang F, Perrotin C, et al. Variable sensitivity and specificity of TTF-1 antibodies in lung metastatic adenocarcinoma of colorectal origin. Mod Pathol. 2005; 18(10):1371-1376.

(41.) Penman D, Downie I, Roberts F. Positive immunostaining for thyroid transcription factor-1 in primary and metastatic colonic adenocarcinoma: a note of caution. J Clin Pathol. 2006; 59(6):663-664.

(42.) Bishop JA, Sharma R, Illei PB. Napsin A and thyroid transcription factor-1 expression in carcinomas of the lung, breast, pancreas, colon, kidney, thyroid, and malignant mesothelioma. Hum Pathol. 2010; 41(1):20-25.

(43.) Cimino-Mathews A, Sharma R, Netto GJ. Diagnostic use of PAX8, CAIX, TTF-1, and TGB in metastatic renal cell carcinoma of the thyroid. Am J Surg Pathol. 2011; 35(5):757-761.

(44.) Ordonez NG. Value of thyroid transcription factor-1, E-cadherin, BG8, WT1, and CD44S immunostaining in distinguishing epithelial pleural mesothelioma from pulmonary and nonpulmonary adenocarcinoma. Am J Surg Pathol. 2000; 24(4):598-606.

(45.) Oliveira AM, Tazelaar HD, Myers JL, Erickson LA, Lloyd RV. Thyroid transcription factor-1 distinguishes metastatic pulmonary from well-differentiated neuroendocrine tumors of other sites. Am J Surg Pathol. 2001; 25(6):815-819.

(46.) Nonaka D, Tang Y, Chiriboga L, Rivera M, Ghossein R. Diagnostic utility of thyroid transcription factors Pax8 and TTF-2 (FoxE1) in thyroid epithelial neoplasms. Mod Pathol. 2008; 21(2):192-200.

(47.) Pasca di Magliano M, Di Lauro R, Zannini M. Pax8 has a key role in thyroid cell differentiation. Proc Natl Acad Sci U S A. 2000; 97(24):13144-13149.

(48.) Chi N, Epstein JA. Getting your Pax straight: Pax proteins in development and disease. Trends Genet. 2002; 18(1):41-47.

(49.) Plachov D, Chowdhury K, Walther C, Simon D, Guenet JL, Gruss P. Pax8, a murine paired box gene expressed in the developing excretory system and thyroid gland. Development. 1990; 110(2):643-651.

(50.) Poleev A, Fickenscher H, Mundlos S, et al. PAX8, a human paired box gene: isolation and expression in developing thyroid, kidney and Wilms' tumors. Development. 1992; 116(3):611-623.

(51.) Kang HC, Ohmori M, Harii N, Endo T, Onaya T. Pax-8 is essential for regulation of the thyroglobulin gene by transforming growth factor-beta1. Endocrinology. 2001; 142(1):267-275.

(52.) Trueba SS, Auge J, Mattei G, et al. PAX8, TITF1, and FOXE1 gene expression patterns during human development: new insights into human thyroid development and thyroid dysgenesis-associated malformations. J Clin Endocrinol Metab. 2005; 90(1):455-462.

(53.) Laury AR, Perets R, Piao H, et al. A comprehensive analysis of PAX8 expression in human epithelial tumors. Am J Surg Pathol. 2011; 35(6):816-826.

(54.) Tacha D, Zhou D, Cheng L. Expression of PAX8 in normal and neoplastic tissues: a comprehensive immunohistochemical study. Appl immunohistochem Mol Morphol. 2011; 19(4):293-299.

(55.) Tong GX, Devaraj K, Hamele-Bena D, et al. Pax8: a marker for carcinoma of Mullerian origin in serous effusions. Diagn Cytopathol. 2011; 39(8):567-574.

(56.) Bishop JA, Sharma R, Westra WH. PAX8 immunostaining of anaplastic thyroid carcinoma: a reliable means of discerning thyroid origin for undifferentiated tumors of the head and neck. Hum Pathol. 2011; 42(12):1873-1877.

(57.) Sangoi AR, Karamchandani J, Kim J, Pai RK, McKenney JK. The use of immunohistochemistry in the diagnosis of metastatic clear cell renal cell carcinoma: a review of PAX-8, PAX-2, hKIM-1, RCCma, and CD10. Adv Anat Pathol. 2010; 17(6):377-393.

(58.) Chernock RD, El-Mofty SK, Becker N, Lewis JS Jr. Napsin A expression in anaplastic, poorly differentiated, and micropapillary pattern thyroid carcinomas. Am I Surg Pathol. 2013; 37(8):1215-1222.

(59.) Bowen NJ, Logani S, Dickerson EB, et al. Emerging roles for PAX8 in ovarian cancer and endosalpingeal development. Gynecol Oncol. 2007; 104(2): 331-337.

(60.) Kobel M, Kalloger SE, Boyd N, et al. Ovarian carcinoma subtypes are different diseases: implications for biomarker studies. PLoS Med. 2008; 5(12): e232.

(61.) Nonaka D, Chiriboga L, Soslow RA. Expression of Pax8 as a useful marker in distinguishing ovarian carcinomas from mammary carcinomas. Am J Surg Pathol. 2008; 32(10):1566-1571.

(62.) Tong GX, Weeden EM, Hamele-Bena D, et al. Expression of PAX8 in nephrogenic adenoma and clear cell adenocarcinoma of the lower urinary tract: evidence of related histogenesis? Am J Surg Pathol. 2008; 32(9):1380-1387.

(63.) Tong GX, Yu WM, Beaubier TN, et al. Expression of PAX8 in normal and neoplastic renal tissues: an immunohistochemical study. Mod Pathol. 2009; 22(9): 1218-1227.

(64.) Ozcan A, de la Roza G, Ro JY, Shen SS, Truong LD. PAX2 and PAX8 expression in primary and metastatic renal tumors: a comprehensive comparison. Arch Pathol Lab Med. 2012; 136(12):1541-1551.

(65.) Ozcan A, Liles N, Coffey D, Shen SS, Truong LD. PAX2 and PAX8 expression in primary and metastatic mullerian epithelial tumors: a comprehensive comparison. Am I Surg Pathol. 2011; 35(12):1837-1847.

(66.) Long KB, Srivastava A, Hirsch MS, et al. PAX8 expression in well-differentiated pancreatic endocrine tumors: correlation with clinicopathologic features and comparison with gastrointestinal and pulmonary carcinoid tumors. Am J Surg Pathol. 2010; 34(5):723-729.

(67.) Sangoi AR, Ohgami RS, Pai RK, Beck AH, McKenney JK, Pai RK. PAX8 expression reliably distinguishes pancreatic well-differentiated neuro-endocrine tumors from ileal and pulmonary well-differentiated neuroendocrine tumors and pancreatic acinar cell carcinoma. Mod Pathol. 2011; 24(3):412-424.

(68.) Liu H, Shi J, Wang HL, Lin F. Identification of effective immunohisto-chemical panels for distinguishing pancreatic endocrine neoplasms from neuroendocrine neoplasms of other organs [CAP poster 24]. Arch Pathol Lab Med. 2012; 136(9):1011.

(69.) Schmitt AC, Cohen C, Siddiqui MT. Paired box gene 8, HBME-1, and cytokeratin 19 expression in preoperative fine-needle aspiration of papillary thyroid carcinoma: diagnostic utility. Cancer Cytopathol. 2010; 118(4):196-202.

(70.) Gudmundsson J, Sulem P, Gudbjartsson DF, et al. Common variants on 9q22.33 and 14q13.3 predispose to thyroid cancer in European populations. Nat Genet. 2009; 41(4):460-464.

(71.) Francis-Lang H, Price M, Polycarpou-Schwarz M, Di Lauro R. Cell-type-specific expression of the rat thyroperoxidase promoter indicates common mechanisms for thyroid-specific gene expression. Mol Cell Biol. 1992; 12(2):576-588.

(72.) Ortiz L, Zannini M, Di Lauro R, Santisteban P. Transcriptional control of the forkhead thyroid transcription factor TTF-2 by thyrotropin, insulin, and insulin-like growth factor I. J Biol Chem. 1997; 272(37):23334-23339.

(73.) Dathan N, Parlato R, Rosica A, De Felice M, Di Lauro R. Distribution of the titf2/foxe1 gene product is consistent with an important role in the development of foregut endoderm, palate, and hair. Dev Dyn. 2002; 224(4): 450-456.

(74.) Zaret KS, Carroll JS. Pioneer transcription factors: establishing competence for gene expression. Genes Dev. 2011; 25(21):2227-2241.

(75.) Cuesta I, Zaret KS, Santisteban P. The forkhead factor FoxE1 binds to the thyroperoxidase promoter during thyroid cell differentiation and modifies compacted chromatin structure. Mol Cell Biol. 2007; 27(20):7302-7314.

(76.) Lloyd RV, Osamura RY. Transcription factors in normal and neo-plastic pituitary tissues. Microsc Res Tech. 1997; 39(2):168-181.

(77.) Sequeira M, Al-Khafaji F, Park S, et al: Production and application of polyclonal antibody to human thyroid transcription factor 2 reveals thyroid transcription factor 2 protein expression in adult thyroid and hair follicles and prepubertal testis. Thyroid. 2003; 13(10):927-932.

(78.) Hishinuma A, Ohmika N, Namatame T, Ieiri T. TTF-2 stimulates expression of 17 genes, including one novel thyroid-specific gene which might be involved in thyroid development. Mol Cell Biol. 2004; 221(1-2):33-46.

(79.) Perrone L, Pasca di Magliano M, Zannini M, Di Lauro R. The thyroid transcription factor 2 (TTF-2) is a promoter-specific DNA-binding independent transcriptional repressor. Biochem Biophys Res Commun. 2000; 275(1):203-208.

(80.) Zannini M, Avantaggiato V, Biffali E, et al. TTF-2, a new forkhead protein, shows a temporal expression in the developing thyroid which is consistent with a role in controlling the onset of differentiation. EMBO J. 1997; 16(11):3185-3197.

(81.) Castanet M, Polak M. Spectrum of human Foxe1/TTF2 mutations. Horm Res Paediatr. 2010; 73(6):423-429.

(82.) Castanet M, Park SM, Smith A, et al. A novel loss-of-function mutation in TTF-2 is associated with congenital hypothyroidism, thyroid agenesis and cleft palate. Hum Mol Genet. 2002; 11(17):2051-2059.

(83.) Matoso A, Easley SE, Mangray S, Jacob R, DeLellis RA. Spindle cell foci in the thyroid gland: an immunohistochemical analysis. Appl Immunohistochem Mol Morphol. 2011; 19(5):400-407.

(84.) Berge-Lefranc JL, Cartouzou G, De Micco C, Fragu P, Lissitzky S. Quantification of thyroglobulin messenger RNA by in situ hybridization in differentiated thyroid cancers: difference between well-differentiated and moderately differentiated histologic types. Cancer. 1985; 56(2):345-350.

(85.) Asioli S, Erickson LA, Righi A, et al. Poorly differentiated carcinoma of the thyroid: validation of the Turin proposal and analysis of IMP3 expression. Mod Pathol. 2010; 23(9):1269-1278.

(86.) Harach HR, Franssila KO. Thyroglobulin immunostaining in follicular thyroid carcinoma: relationship to the degree of differentiation and cell type. Histopathology. 1988; 13(1):43-54.

(87.) Carcangiu ML, Steeper T, Zampi G, Rosai J. Anaplastic thyroid carcinoma: a study of 70 cases. Am J Clin Pathol. 1985; 83(2):135-158.

(88.) Albores-Saavedra J, Nadji M, Civantos F, Morales AR. Thyroglobulin in carcinoma of the thyroid: an immunohistochemical study. Hum Pathol. 1983; 14(1):62-66.

(89.) Logmans SC, Jobsis AC. Thyroid-associated antigens in routinely embedded carcinomas: possibilities and limitations studied in 116 cases. Cancer. 1984; 54(2):274-279.

(90.) Heffess CS, Wenig BM, Thompson LD. Metastatic renal cell carcinoma to the thyroid gland: a clinicopathologic study of 36 cases. Cancer. 2002; 95(9): 1869-1878.

(91.) Sathiyamoorthy S, Maleki Z. Cytomorphologic overlap of differentiated thyroid carcinoma and lung adenocarcinoma and diagnostic value of TTF-1 and TGB on cytologic material [published online ahead of print May 2, 2013.] Diagn Cytopathol. doi:10.1002/dc.22997.

(92.) Liu H, Lin F, DeLellis RA. Thyroid and parathyroid gland. In: Lin F, Prichard JW, Liu H, Wilkerson M, Scheurch C, eds. Handbook of Practical Immunohistochemistry: Frequently Asked Questions. New York, NY: Springer; 2011:137-158.

(93.) Rezk S, Khan A. Role of immunohistochemistry in the diagnosis and progression of follicular epithelium-derived thyroid carcinoma. Appl Immunohistochem Mol Morphol. 2005; 13(3):256-264.

(94.) Fischer S, Asa SL. Application of immunohistochemistry to thyroid neoplasms. Arch Pathol Lab Med. 2008; 132(3):359-372.

(95.) Saleh HA, Jin B, Barnwell J, Alzohaili O. Utility of immunohisto-chemical markers in differentiating benign from malignant follicular-derived thyroid nodules. Diagn Pathol. 2010; 5:9.

(96.) Chiu CG, Strugnell SS, Griffith OL, et al. Diagnostic utility of galectin-3 in thyroid cancer. Am J Pathol. 2010; 176(5):2067-2081.

(97.) Ruggeri RM, Campenm A, Baldari S, Trimarchi F, Trovato M. What is new on thyroid cancer biomarkers. Biomark Insights. 2008; 3:237-252.

(98.) de Matos LL, Del Giglio AB, Matsubayashi CO, de Lima Farah M, Del Giglio A, da Silva Pinhal MA. Expression of CK-19, galectin-3 and HBME-1 in the differentiation of thyroid lesions: systematic review and diagnostic meta-analysis. Diagn Pathol. 2012; 7:97.

(99.) Sobrinho-Simoes M, Eloy C, Magalhaes J, Lobo C, Amaro T. Follicular thyroid carcinoma. Mod Pathol. 2011; 24(suppl 2):S10-S18.

(100.) Nakamura N, Erickson LA, Jin L, et al. Immunohistochemical separation of follicular variant of papillary thyroid carcinoma from follicular adenoma. Endocr Pathol. 2006; 17(3):213-223.

(101.) Paunovic I, Isic T, Havelka M, Tatic S, Cvejic D, Savin S. Combined immunohistochemistry for thyroid peroxidase, galectin-3, CK19 and HBME-1 in differential diagnosis of thyroid tumors. APMIS. 2012; 120(5):368-379.

(102.) Park YJ, Kwak SH, Kim DC, et al. Diagnostic value of galectin-3, HBME-1, cytokeratin 19, high molecular weight cytokeratin, cyclin D1 and p27(kip1) in the differential diagnosis of thyroid nodules. J Korean Med Sci. 2007; 22(4):621-628.

(103.) Savin S, Cvejic D, Isic T, Paunovic I, Tatic S, Havelka M. Thyroid peroxidase and galectin-3 immunostaining in differentiated thyroid carcinoma with clinicopathologic correlation. Hum Pathol. 2008; 39(11):1656-1663.

(104.) Volante M, Bozzalla-Cassione F, DePompa R, et al. Galectin-3 and HBME-1 expression in oncocytic cell tumors of the thyroid. Virchows Arch. 2004; 445(2):183-188.

(105.) Barut F, Onak Kandemir N, Bektas S, et al. Universal markers of thyroid malignancies: galectin-3, HBME-1, and cytokeratin-19. Endocr Pathol. 2010; 21(2):80-89.

(106.) Saggiorato E, De Pompa R, Volante M, et al. Characterization of thyroid 'follicular neoplasms' in fine-needle aspiration cytological specimens using a panel of immunohistochemical markers: a proposal for clinical application. Endocr Relat Cancer. 2005; 12(2):305-317.

(107.) Rossi ED, Raffaelli M, Mule? A, et al. Simultaneous immunohistochemical expression of HBME-1 and galectin-3 differentiates papillary carcinomas from hyperfunctioning lesions of the thyroid. Histopathology. 2006; 48(7):795-800.

(108.) Prasad ML, Pellegata NS, Huang Y, Nagaraja Hn, de la Chapelle A, Kloos RT. Galectin-3, fibronectin-1, CITED-1, HBME1 and cytokeratin-19 immunohisto-chemistry is useful for the differential diagnosis of thyroid tumors. Mod Pathol. 2005; 18(1):48-57.

(109.) Liu YY, Morreau H, Kievit J, Romijn JA, Carrasco N, Smit JW. Combined immunostaining with galectin-3, fibronectin-1, CITED-1, Hector Battifora mesothelial-1, cytokeratin-19, peroxisome proliferator-activated receptor-{gamma}, and sodium/iodide symporter antibodies for the differential diagnosis of non-medullary thyroid carcinoma. Eur J Endocrinol. 2008; 158(3):375-384.

(110.) de Matos PS, Ferreira AP, de Oliveira Facuri F, Assumpcao LV, Metze K, Ward LS. Usefulness of HBME-1, cytokeratin 19 and galectin-3 immunostaining in the diagnosis of thyroid malignancy. Histopathology. 2005; 47(4):391-401.

(111.) Beesley MF, McLaren KM. Cytokeratin 19 and galectin-3 immunohisto-chemistry in the differential diagnosis of solitary thyroid nodules. Histopathology. 2002; 41(3):236-243.

(112.) Scognamiglio T, Hyjek E, Kao J, Chen YT. Diagnostic usefulness of HBME1, galectin-3, CK19, and CITED1 and evaluation of their expression in encapsulated lesions with questionable features of papillary thyroid carcinoma. Am J Clin Pathol. 2006; 126(5):700-708.

(113.) Liu H, Lin F. TROP-2 is a potential novel immunomarker for identification of papillary thyroid carcinomas. Accepted for presentation at: USCAP Annual Meeting; March 1-8, 2014; San Diego, CA.

(114.) Barroeta JE, Baloch ZW, Lal P, Pasha TL, Zhang PJ, LiVolsi VA. Diagnostic value of differential expression of CK19, Galectin-3, HBME-1, ERK, RET, and p16 in benign and malignant follicular-derived lesions of the thyroid: an immunohisto-chemical tissue microarray analysis. Endocr Pathol. 2006; 17(3):225-234.

(115.) Nasr MR, Mukhopadhyay S, Zhang S, Katzenstein AL. Immunohisto-chemical markers in diagnosis of papillary thyroid carcinoma: utility of HBME1 combined with CK19 immunostaining. Mod Pathol. 2006; 19(12):1631-1637.

(116.) Mai KT, Bokhary R, Yazdi HM, Thomas J, Commons AS. Reduced HBME1 immunoreactivity of papillary thyroid carcinoma and papillary thyroid carcinoma-related neoplastic lesions with Hurthle cell and/or apocrine-like changes. Histopathology. 2002; 40(2):133-142.

(117.) Volante M, Bozzalla-Cassione F, Orlandi F, Papotti M. Diagnostic role of galectin-3 in follicular thyroid tumors. Virchows Arch. 2004; 444(4):309-312.

(118.) Cheung CC, Ezzat S, Freeman JL, Rosen IB, Asa SL. Immunohistochemical diagnosis of papillary thyroid carcinoma. Mod Pathol. 2001; 14(4):338-342.

(119.) Lenggenhager D, Maggio EM, Moch H, Rossle M. HBME-1 expression in hyalinizing trabecular tumours of the thyroid gland. Histopathology. 2013; 62(7): 1092-1097.

(120.) Rossi ED, Straccia P, Palumbo M, et al. Diagnostic and prognostic role of HBME-1, galectin-3, and b-catenin in poorly differentiated and anaplastic thyroid carcinomas. Appl Immunohistochem Mol Morphol. 2013; 21(3):237-241.

(121.) Nechifor-Boila A, Borda A, Sassolas G, et al. Immunohistochemical markers in the diagnosis of papillary thyroid carcinomas: the promising role of combined immunostaining using HBME-1 and CD56. Pathol Res Pract. 2013; 209(9):585-592.

(122.) Torregrossa L, Faviana P, Camacci T, et al. Galectin-3 is highly expressed in nonencapsulated papillary thyroid carcinoma but weakly expressed in encapsulated type; comparison with Hector Battifora mesothelial cell 1 immunoreactivity. Hum Pathol. 2007; 38(10)1482-1488.

(123.) Mase T, Funahashi H, Koshikawa T, et al. HBME-1 immunostaining in thyroid tumors especially in follicular neoplasm. Endocr J. 2003; 50(2):173-177.

(124.) Papotti M, Rodriguez J, De Pompa R, Bartolazzi A, Rosai J. Galectin-3 and HBME-1 expression in well-differentiated thyroid tumors with follicular architecture of uncertain malignant potential. Mod Pathol. 2005; 18(4):541-546.

(125.) Ito Y, Yoshida H, Tomoda C, et al. HBME-1 expression in follicular tumor of the thyroid: an investigation of whether it can be used as a marker to diagnose follicular carcinoma. Anticancer Res. 2005; 25(1A):179-182.

(126.) Zhu X, Sun T, Lu H, et al. Diagnostic significance of CK19, RET, galectin-3 and HBME-1 expression for papillary thyroid carcinoma. J Clin Pathol. 201063(9): 786-789.

(127.) Wiseman SM, Melck A, Masoudi H, et al. Molecular phenotyping of thyroid tumors identifies a marker panel for differentiated thyroid cancer diagnosis. Ann Surg Oncol. 2008; 15(10):2811-2826.

(128.) Asa SL. The role of immunohistochemical markers in the diagnosis of follicular-patterned lesions of the thyroid. Endocr Pathol. 2005; 16(4):295-309.

(129.) Baloch ZW, Abraham S, Roberts S, LiVolsi VA. Differential expression of cytokeratins in follicular variant of papillary carcinoma: an immunohistochemical study and its diagnostic utility. Hum Pathol. 1999; 30(10):1166-1171.

(130.) Murphy KM, Chen F, Clark DP. Identification of immunohistochemical biomarkers for papillary thyroid carcinoma using gene expression profiling. Hum Pathol. 2008; 39(3):420-426.

(131.) Erkilic S, Aydin A, Kocer NE. Diagnostic utility of cytokeratin 19 expression in multinodular goiter with papillary areas and papillary carcinoma of thyroid. Endocr Pathol. 2002; 13(3):207-211.

(132.) Sahoo S, Hoda SA, Rosai J, DeLellis RA. Cytokeratin 19 immunoreactivity in the diagnosis of papillary thyroid carcinoma: a note of caution. Am J Clin Pathol. 2001; 116(5):696-702.

(133.) Song Q, Wang D, Lou Y, et al. Diagnostic significance of CK19, TG, Ki67 and galectin-3 expression for papillary thyroid carcinoma in the northeastern region of China. Diagn Pathol. 2011; 6:126.

(134.) Bose D, Das RN, Chatterjee U, Banerjee U. Cytokeratin 19 immunoreactivity in the diagnosis of papillary thyroid carcinoma. Indian J Med Paediatr Oncol. 2012; 33(2):107-111.

(135.) Casey MB, Lohse CM, Lloyd RV. Distinction between papillary thyroid hyperplasia and papillary thyroid carcinoma by immunohistochemical staining for cytokeratin 19, galectin-3, and HBME-1. Endocr Pathol. 2003; 14(1):55-60.

(136.) Gong HC, Honjo Y, Nangia-Makker P, et al. The NH2 terminus of galectin-3 governs cellular compartmentalization and functions in cancer cells. Cancer Res. 1999; 59(24):6239-6245.

(137.) Cowles EA, Agrwal N, Anderson RL, Wang JL. Carbohydrate-binding protein 35. Isoelectric points of the polypeptide and a phosphorylated derivative. J Biol Chem. 1990; 265(29):17706-17712.

(138.) Weber KB, Shroyer KR, Heinz DE, Nawaz S, Said MS, Haugen BR. The use of a combination of galectin-3 and thyroid peroxidase for the diagnosis and prognosis of thyroid cancer. Am J Clin Pathol. 2004; 122(4):524-531.

(139.) Saggiorato E, Aversa S, Deandreis D, et al. Galectin-3: presurgical marker of thyroid follicular epithelial cell-derived carcinomas. J Endocrinol Invest. 2004; 27(4):311-317.

(140.) Orlandi F, Saggiorato E, Pivano G, et al. Galectin-3 is a presurgical marker of human thyroid carcinoma. Cancer Res. 1998; 58(14):3015-3020.

(141.) Saggiorato E, Cappia S, De Giuli P, et al. Galectin-3 as a presurgical immunocytodiagnostic marker of minimally invasive follicular thyroid carcinoma. J Clin Endocrinol Metab. 2001; 86(11):5152-5158.

(142.) Bartolazzi A, Orlandi F, Saggiorato E, et al; Italian Thyroid Cancer Study Group (ITCSG). Galectin-3-expression analysis in the surgical selection of follicular thyroid nodules with indeterminate fine-needle aspiration cytology: a prospective multicentre study. Lancet Oncol. 2008; 9(6):543-549.

(143.) Gaffney RL, Carney JA, Sebo TJ, et al. Galectin-3 expression in hyalinizing trabecular tumors of the thyroid gland. Am J Surg Pathol. 2003; 27(4):494-498.

(144.) Coli A, Bigotti G, Zucchetti F, Negro F, Massi G. Galectin-3, a marker of well-differentiated thyroid carcinoma, is expressed in thyroid nodules with cytological atypia. Histopathology. 2002; 40(1):80-87.

(145.) Nascimento MC, Bisi H, Alves VA, Longatto-Filho A, Kanamura CT, Medeiros-Neto G. Differential reactivity for galectin-3 in Hurthle cell adenomas and carcinomas. Endocr Pathol. 2001; 12(3):275-279.

(146.) Bartolazzi A, Gasbarri A, Papotti M, et al; Thyroid Cancer Study Group. Application of an immunodiagnostic method for improving preoperative diagnosis of nodular thyroid lesions. Lancet. 2001; 357(9269):1644-1650.

(147.) Wenqi D, Li W, Shanshan C, et al. EpCAM is overexpressed in gastric cancer and its downregulation suppresses proliferation of gastric cancer. J Cancer Res Clin Oncol. 2009; 135(9):1277-1285.

(148.) Cubas R, Li M, Chen C, Yao Q. Trop2: a possible therapeutic target for late stage epithelial carcinomas. Biochim Biophys Acta. 2009; 1796(2):309-314.

(149.) Ohmachi T, Tanaka F, Mimori K, Inoue H, Yanaga K, Mori M. Clinical significance of TROP2 expression in colorectal cancer. Clin Cancer Res. 2006; 12(10):3057-3063.

(150.) Fong D, Spizzo G, Gostner JM, et al. TROP2: a novel prognostic marker in squamous cell carcinoma of the oral cavity. Mod Pathol. 2008; 21(2):186-191.

(151.) Lipinski M, Parks DR, Rouse RV, Herzenberg LA. Human trophoblastcellsurface antigens defined by monoclonal antibodies. Proc Natl Acad Sci USA. 1981; 78(8):5147-5150.

(152.) Alberti S, Miotti S, Stella M, et al. Biochemical characterization of Trop-2, a cell surface molecule expressed by human carcinomas: formal proof that the monoclonal antibodies T16 and MOv-16 recognize Trop-2. Hybridoma. 1992; 11(5):539-545.

(153.) Sukhthankar M, Alberti S, Baek SJ. (-)-Epigallocatechin-3-gallate (EGCG) post-transcriptionally and post-translationally suppresses the cell proliferative protein TROP2 in human colorectal cancer cells. Anticancer Res. 2010; 30(7):

2497-2503.

(154.) Muhlmann G, Spizzo G, Gostner J, et al. TROP2 expression as prognostic marker for gastric carcinoma. J Clin Pathol. 2009; 62(2):152-158.

(155.) Fong D, Moser P, Krammel C, et al. High expression of TROP2 correlates with poor prognosis in pancreatic cancer. Br J Cancer. 2008; 99(8):1290-1295.

(156.) Pak MG, Shin DH, Lee Ch, Lee MK. Significance of EpCAM and TROP2 expression in non-small cell lung cancer. World J Surg Oncol. 2012; 10:53.

(157.) Fornaro M, Dell'Arciprete R, Stella M, et al. Cloning of the gene encoding Trop-2, a cell-surface glycoprotein expressed by human carcinomas. Int J Cancer. 1995; 62(5):610-618.

(158.) Stepan LP, Trueblood ES, Hale K, Babcook J, Borges L, Sutherland CL. Expression of Trop2 cell surface glycoprotein in normal and tumor tissues: potential implications as a cancer therapeutic target. J Histochem Cytochem. 2011; 59(7):701-710.

(159.) Kobayashi H, Minami Y, Anami Y, et al. Expression of the GA733 gene family and its relationship to prognosis in pulmonary adenocarcinoma. Virchows Arch. 2010; 457(1):69-76.

(160.) Ning S, Guo S, Xie J, Xu Y, Lu X, Chen Y. TROP2 correlates with microvessel density and poor prognosis in hilar cholangiocarcinoma. J Gastrointest Surg. 2013; 17(2):360-368.

(161.) Bignotti E, Todeschini P, Calza S, et al. Trop-2 overexpression as an independent marker for poor overall survival in ovarian carcinoma patients. Eur J Cancer. 2010; 46(5):944-953.

(162.) Fang YJ, Lu ZH, Wang GQ, et al. Elevated expressions of MMP7, TROP2, and survivin are associated with survival, disease recurrence, and liver metastasis of colon cancer. Int J Colorectal Dis. 2009; 24(8):875-884.

(163.) Trerotola M, Cantanelli P, Guerra E, et al. Upregulation of Trop-2 quantitatively stimulates human cancer growth. Oncogene. 2013; 32(2):222-233.

(164.) De Micco C, Ruf J, Chrestian MA, Gros N, Henry JF, Carayon P. Immunohistochemical study of thyroid peroxidase in normal, hyperplastic, and neoplastic human thyroid tissues. Cancer. 1991; 67(12):3036-3041.

(165.) Yousaf U, Christensen LH, Rasmussen AK, et al. Immunohistochemical staining for thyroid peroxidase (TPO) of needle core biopsies in the diagnosis of scintigraphically cold thyroid nodules. Clin Endocrinol (Oxf). 2008; 68(6):996-1001.

(166.) Christensen L, Blichert-Toft M, Brandt M, et al. Thyroperoxidase (TPO) immunostaining of the solitary cold thyroid nodule. Clin Endocrinol (Oxf). 2000; 53(2):161-169.

(167.) De Micco C, Vasko V, Garcia S, Zoro P, Denizot A, Henry JF. Fine-needle aspiration of thyroid follicular neoplasm: diagnostic use of thyroid peroxidase immunocytochemistry with monoclonal antibody 47. Surgery. 1994; 116(6): 1031-1035.

(168.) de Micco C, Savchenko V, Giorgi R, Sebag F, Henry JF. Utility of malignancy markers in fine-needle aspiration cytology of thyroid nodules: comparison of Hector Battifora mesothelial antigen-1, thyroid peroxidase and dipeptidyl aminopeptidase IV. Br J Cancer. 2008; 98(4):818-823.

(169.) Savin S, Cvejic D, Isic T, et al. Thyroid peroxidase immunohistochemistry in differential diagnosis of thyroid tumors. Endocr Pathol. 2006; 17(1):53-60.

(170.) Scheumman GF, Hoang-Vu C, Cetin Y, et al. Clinical significance of Ecadherin as a prognostic marker in thyroid carcinomas. J Clin Endocrinol Metab. 1995; 80(7):2168-2172.

(171.) von Wasielewski R, Rhein A, Werner M, et al. Immunohistochemical detection of E-cadherin in differentiated thyroid carcinomas correlates with clinical outcome. Cancer Res. 1997; 57(12):2501-2507.

(172.) Brabant G, Hoang-Vu C, Cetin Y, et al. E-cadherin: a differentiation marker in thyroid malignancies. Cancer Res. 1993; 53(20):4987-4993.

(173.) Rocha AS, Soares P, Fonseca E, Cameselle-Teijeiro J, Oliveira MC, Sobrinho-Simoes M. E-cadherin loss rather than beta-catenin alterations is a common feature of poorly differentiated thyroid carcinomas. Histopathology. 2003; 42(6):580-587.

(174.) Van Aken E, De Wever O, Correia da Rocha AS, Mareel M. Defective Ecadherin/catenin complexes in human cancer. Virchows Arch. 2001; 439(6):725751.

(175.) Kato N, Tsuchiya T, Tamura G, Motoyama T. E-cadherin expression in follicular carcinoma of the thyroid. Pathol Int. 2002; 52(1):13-18.

(176.) Ryu S, Jimi S, Takebayashi S. Thyroid carcinoma distinctively expresses intracellular fibronectin in vivo. Cancer Lett. 1997; 121(2):189-193.

(177.) Scarpino S, Stoppacciaro A, Pellegrini C, et al. Expression of EDA/EDB isoforms of fibronectin in papillary carcinoma of the thyroid. J Pathol. 1999; 188(2):163-167.

(178.) Takano T, Miyauchi A, Yokozawa T, et al. Preoperative diagnosis of thyroid papillary and anaplastic carcinomas by real-time quantitative reverse transcription-polymerase chain reaction of oncofetal fibronectin messenger RNA. Cancer Res. 1999; 59(18):4542-4545.

(179.) Giannini R, Faviana P, Cavinato T, et al. Galectin-3 and oncofetalfibronectin expression in thyroid neoplasia as assessed by reverse transcriptionpolymerase chain reaction and immunochemistry in cytologic and pathologic specimens. Thyroid. 2003; 13(8):765-770.

(180.) Huang Y, Prasad M, Lemon WJ, et al. Gene expression in papillary thyroid carcinoma reveals highly consistent profiles. Proc Natl Acad Sci USA. 2001; 98(26):15044-15049.

(181.) Aruffo A, Stamenkovic I, Melnick M, Underhill CB, Seed B. CD44 is the principal cell surface receptor for hyaluronate. Cell. 1990; 61(7):1303-1313.

(182.) Rudzki Z, Jothy S. CD44 and the adhesion of neoplastic cells. Mol Pathol. 1997; 50(2):57-71.

(183.) Xin Y, Grace A, Gallagher MM, Curran BT, Leader MB, Kay EW. CD44V6 in gastric carcinoma: a marker of tumor progression. Appl immunohistochem Mol Morphol. 2001; 9(2):138-142.

(184.) Bendardaf R, Lamlum H, Ristamaki R, Pyrhonen S. CD44 variant 6 expression predicts response to treatment in advanced colorectal cancer. Oncol Rep. 2004; 11(1):41-45.

(185.) Rodrigo JP, Dominguez F, Alvarez C, Herrero A, Suarez C. Expression of E-cadherin, CD44s, and CD44v6 in laryngeal and pharyngeal carcinomas. Am J Otolaryngol. 2003; 24(6):384-389.

(186.) Kurozumi K, Nakao K, Nishida T, Nakahara M, Ogino N, Tsujimoto M. Significance of biologic aggressiveness and proliferating activity in papillary thyroid carcinoma. World I Surg. 1998; 22(12):1237-1242.

(187.) Gu J, Daa T, Kashima K, Yokoyama S, Nakayama I, Noguchi S. Expression of splice variants of CD44 in thyroid neoplasms derived from follicular cells. Pathol int. 1998; 48(3):184-190.

(188.) Figge J, del Rosario AD, Gerasimov G, et al. Preferential expression of the cell adhesion molecule CD44 in papillary thyroid carcinoma. Exp Mol Pathol. 1994; 61(3):203-211.

(189.) Kim J, Cho H, Rhee B, Kim HY. Expression of CD44 and cyclin D1 in fine needle aspiration cytology of papillary thyroid carcinoma. Acta Cytol. 2002; 46(4):679-683.

(190.) Gasbarri A, Martegani MP, Del Prete F, Lucante T, Natali PG, Bartolazzi A. Galectin-3 and CD44v6 isoforms in the preoperative evaluation of thyroid nodules. J Clin Oncol. 1999; 17(11):3494-3502.

(191.) Maruta J, Hashimoto H, Yamashita H, Yamashita H, Noguchi S. Immunostaining of galectin-3 and CD44v6 using fine-needle aspiration for distinguishing follicular carcinoma from adenoma. Diagn Cytopathol. 2004; 31(6):392-396.

(192.) Catzavelos C, Bhattacharya N, Ung YC, et al. Decreased levels of the cell cycle inhibitor p27Kip1 protein: prognostic implications in primary breast cancer. Nat Med. 1997; 3(2):227-230.

(193.) Fredersdorf S, Burns J, Milne AM, et al. High level expression of p27(kip1) and cyclin D1 in some human breast cancer cells: inverse correlation between the expression of p27(kip1) and degree of malignancy in human breast and colorectal cancers. Proc Natl Acad Sci USA. 1997; 94(12):6380-6385.

(194.) Loda M, Cukor B, Tam SW, et al. Increased proteasome-dependent degradation of the cyclin-dependent kinase inhibitor p27 in aggressive colorectal carcinomas. Nat Med. 1997; 3(2):231-234.

(195.) Singh SP, Lipman J, Goldman H, et al. Loss or altered subcellular localization of p27 in Barrett's associated adenocarcinoma. Cancer Res. 1998; 58(8):1730-1735.

(196.) Tsihlias J, Kapusta LR, DeBoer G, et al. Loss of cyclin-dependent kinase inhibitor p27kip1 is a novel prognostic factor in localized human prostate adenocarcinoma. Cancer Res. 1998; 58(3):542-548.

(197.) Erickson LA, Jin L, Wollan PC, Thompson GB, van Heerden J, Lloyd RV. Expression of p27kip1 and Ki-67 in benign and malignant thyroid tumors. Mod Pathol. 1998; 11(2):169-174.

(198.) Resnick MB, Schacter P, Finkelstein Y, Kellner Y, Cohen O. Immunohistochemical analysis of p27/kip1 expression in thyroid carcinoma. Mod Pathol. 1998; 11(8):735-739.

(199.) Erickson LA, Yousef OM, Jin L, Lohse CM, Pankratz VS, Lloyd RV. p27kip1 expression distinguishes papillary hyperplasia in Graves' disease from papillary thyroid carcinoma. Mod Pathol. 2000; 13(9):1014-1019.

(200.) Tallini G, Garcia-Rostan G, Herrero A, et al. Downregulation of p27KIP1 and Ki67/Mib1 labeling index support the classification of thyroid carcinoma into prognostically relevant categories. Am J Surg Pathol. 1999; 23(6):678-685.

(201.) Wang S, Wuu J, Savas L, Patwardhan N, Khan A. The role of cell cycle regulatory proteins, cyclin D1, cyclin E, and p27 in thyroid carcinogenesis. Hum Pathol. 1998; 29(11):1304-1309.

(202.) Yang K, Hitomi M, Stacey DW. Variations in cyclin D1 levels through the cell cycle determine the proliferative fate of a cell. Cell Div. 2006; 1:32.

(203.) Buckley MF, Sweeney KJ, Hamilton JA, et al. Expression and amplification of cyclin genes in human breast cancer. Oncogene. 1993; 8(8):2127-2133.

(204.) Jiang W, Kahn SM, Tomita N, Zhang YJ, Lu SH, Weinstein IB. Amplification and expression of the human cyclin D gene in esophageal cancer. Cancer Res. 1992; 52(10):2980-2983.

(205.) Michalides R, van Veelen N, Hart A, Loftus B, Wientjens E, Balm A. Overexpression of cyclin D1 correlates with recurrence in a group of forty-seven operable squamous cell carcinomas of the head and neck. Cancer Res. 1995; 55(5):975-978.

(206.) Lazzereschi D, Sambuco L, Carnovale Scalzo C, et al. Cyclin D1 and cyclin E expression in malignant thyroid cells and in human thyroid carcinomas. int! Cancer. 1998; 76(6):806-811.

(207.) Brzezianska E, Cyniak-Magierska A, Sporny S, Pastuszak-Lewandoska D, Lewrnski A. Assessment of cyclin D1 gene expression as a prognostic factor in benign and malignant thyroid lesions. Neuro Endocrinol Lett. 2007; 28(4):341350.

(208.) Temmim L, Ebraheem AK, Baker H, Sinowatz F. Cyclin D1 protein expression in human thyroid gland and thyroid cancer. Anat Histol Embryol. 2006; 35(2):125-129.

(209.) Lee SH, Lee JK, Jin SM, et al. Expression of cell-cycle regulators (cyclin D1, cyclin E, p27kip1, p57kip2) in papillary thyroid carcinoma. Otolaryngol Head Neck Surg. 2010; 142(3):332-337.

(210.) Melck A, Masoudi H, Griffith OL, et al. Cell cycle regulators show diagnostic and prognostic utility for differentiated thyroid cancer. Ann Surg Oncol. 2007; 14(12):3403-3411.

(211.) Seybt TP, Ramalingam P, Huang J, Looney SW, Reid MD. Cyclin D1 expression in benign and differentiated malignant tumors of the thyroid gland: diagnostic and biologic implications. Appl Immunohistochem Mol Morphol. 2012; 20(2):124-130.

(212.) Wang S, Lloyd RV, Hutzler MJ, Safran MS, Patwardhan NA, Khan A. The role of cell cycle regulatory protein, cyclin D1, in the progression of thyroid cancer. Mod Pathol. 2000; 13(8):882-887.

(213.) Erickson LA, Jin L, Goellner JR, et al. Pathologic features, proliferative activity, and cyclin D1 expression in Hurthle cell neoplasms of the thyroid. Mod Pathol. 2000; 13(2):186-192.

(214.) Cerrato A, Fulciniti F, Avallone A, Benincasa G, Palombini L, Grieco M. Beta- and gamma-catenin expression in thyroid carcinomas. J Pathol. 1998; 185(3):267-272.

(215.) Dierick H, Bejsovec A. Cellular mechanisms of wingless/Wnt signal transduction. Curr Top Dev Biol. 1999; 43:153-190.

(216.) Miller JR, Hocking AM, Brown JD, Moon RT. Mechanism and function of signal transduction by the Wnt/beta-catenin and Wnt/Ca2+ pathways. Oncogene. 1999; 18(55):7860-7872.

(217.) Peifer M, Polakis P. Wnt signaling in oncogenesis and embryogenesis--a look outside the nucleus. Science. 2000; 287(5458):1606-1609.

(218.) Garcia-Rostan G, Camp RL, Herrero A, Carcangiu ML, Rimm DL, Tallini G. Beta-catenin dysregulation in thyroid neoplasms: down-regulation, aberrant nuclear expression, and CTNNB1 exon 3 mutations are markers for aggressive tumor phenotypes and poor prognosis. Am J Pathol. 2001; 158(3):987-996.

(219.) Rezk S, Brynes RK, Nelson V, et al. beta-Catenin expression in thyroid follicular lesions: potential role in nuclear envelope changes in papillary carcinomas. Endocr Pathol. 2004; 15(4):329-337.

(220.) Ishigaki K, Namba H, Nakashima M, et al. Aberrant localization of betacatenin correlates with overexpression of its target gene in human papillary thyroid cancer. J Clin Endocrinol Metab. 2002; 87(7):3433-3440.

(221.) Pilotti S, Collini P, Del Bo R, Cattoretti G, Pierotti MA, Rilke F. A novel panel of antibodies that segregates immunocytochemically poorly differentiated carcinoma from undifferentiated carcinoma of the thyroid gland. Am J Surg Pathol. 1994; 18(10):1054-1064.

(222.) Lam KY, Lo CY, Chan KW, Wan KY. Insular and anaplastic carcinoma of the thyroid: a 45-year comparative study at a single institution and a review of the significance of p53 and p21. Ann Surg. 2000; 231(3):329-338.

(223.) Evans JJ, Crist HS, Durvesh S, Bruggeman RD, Goldenberg D. A comparative study of cell cycle mediator protein expression patterns in ana-plastic and papillary thyroid carcinoma. Cancer Biol Ther. 2012; 13(9):776-781.

(224.) Quiros RM, Ding HG, Gattuso P, Prinz RA, Xu X. Evidence that one subset of anaplastic thyroid carcinomas are derived from papillary carcinomas due to BRAF and p53 mutations. Cancer. 2005; 103(11):2261-2268.

(225.) Dobashi Y, Sugimura H, Sakamoto A, et al. Stepwise participation of p53 gene mutation during dedifferentiation of human thyroid carcinomas. Diagn Mol Pathol. 1994; 3(1):9-14.

(226.) Donghi R, Longoni A, Pilotti S, Michieli P, Della Porta G, Pierotti MA. Gene p53 mutations are restricted to poorly differentiated and undifferentiated carcinomas of the thyroid gland. J Clin invest. 1993; 91(4):1753-1760.

(227.) Routhier CA, Mochel MC, Lynch K, Dias-Santagata D, Louis DN, Hoang MP. Comparison of 2 monoclonal antibodies for immunohistochemical detection of BRAF V600E mutation in malignant melanoma, pulmonary carcinoma, gastrointestinal carcinoma, thyroid carcinoma, and gliomas. Hum Pathol. 2013; 44(11):2563-2570.

(228.) McKelvie PA, Chan F, Yu Y, et al. The prognostic significance of the BRAFV600E mutation in papillary thyroid carcinoma detected by mutationspecific immunohistochemistry. Pathology. 2013; 45(7):637-644.

(229.) Kim TH, Park YJ, Lim JA, et al. The association of the BRAF(V600E) mutation with prognostic factors and poor clinical outcome in papillary thyroid cancer: a meta-analysis. Cancer. 2012; 118(7):1764-1773.

(230.) Zagzag J, Pollack A, Dultz L, et al. Clinical utility of immunohisto-chemistry for the detection of the BRAF v600e mutation in papillary thyroid carcinoma. Surgery. 2013; 154(6):1199-1205.

(231.) Bullock M, O'Neill C, Chou A, et al. Utilization of a MAB for BRAF(V600E) detection in papillary thyroid carcinoma. Endocr Relat Cancer. 2012; 19(6):779-784.

(232.) Koperek O, Kornauth C, Capper D, et al. Immunohistochemical detection of the BRAF V600E-mutated protein in papillary thyroid carcinoma. Am J Surg Pathol. 2012; 36(6):844-850.

(233.) Capper D, Preusser M, Habel A, et al. Assessment of BRAF V600E mutation status by immunohistochemistry with a mutation-specific monoclonal antibody. Acta Neuropathol. 2011; 122(1):11-19.

(234.) Cameron BR, Berean KW. Cytokeratin subtypes in thyroid tumours: immunohistochemical study with emphasis on the follicular variant of papillary carcinoma. J Otolaryngol. 2003; 32(5):319-322.

Haiyan Liu, MD; Fan Lin, MD, PhD

Accepted for publication March 10, 2014.

From the Department of Laboratory Medicine, Geisinger Medical Center, Danville, Pennsylvania.

The authors have no relevant financial interest in the products or companies described in this article.

Reprints: Haiyan Liu, MD, Department of Laboratory Medicine, MC 01-31, Geisinger Medical Center, 100 N Academy Ave, Danville, PA 17822 (e-mail: hliu1@geisinger.edu).

* References 18, 25, 26, 28, 30, 31, 39, 43-46.

([dagger]) References 95, 104-110, 112, 114-116,118,120-128.

* References 95, 104-110, 112, 114-116, 118, 119, 121, 123-125, 127.

([section]) References 95, 104-108,110-112,114, 115,118,130-135.

([parallel]) References 95, 103-112, 114, 117, 120, 122, 124, 126-128, 130, 133, 138-146.

Caption: Figure 1. A through F, Trophoblastic cell surface antigen 2 (TROP2) staining pattern in thyroid neoplasm and lesions. A, Papillary thyroid carcinoma (PTC), follicular variant. B, Papillary thyroid carcinoma, follicular variant, diffuse (4+) TROP2 staining, membranous pattern. Ninety percent of PTCs on tissue microarray sections express TROP2 in a membranous staining pattern. C, Follicular thyroid carcinoma (FTC). D, Follicular thyroid carcinoma, focal (1+) strong cytoplasmic staining for TROP2. Follicular neoplasms (FTC and follicular adenoma [FA]) showed no TROP2 expression. Only 2 of 51 FAs and 4 of 37 FTCs showed focal (1+) cytoplasmic staining without membranous pattern. E, Focal cystic degeneration in a case of lymphocytic thyroiditis (LTis). F, Focal membranous staining for TROP2 in the lining cells of the cyst. All 20 cases of benign thyroid lesions (10 Ltis, 10 nodular goiter) did not show TROP2 staining (hematoxylin-eosin, original magnifications X400 [A, C] and X200 [E]; original magnifications X400 [B, D] and X200 [F]).

Caption: Figure 2. A through D, Immunohistochemical detection of BRAF expression in a papillary thyroid microcarcinoma and its metastasis in a lymph node, using anti-VE1 antibody. A, Papillary thyroid microcarcinoma. B, Diffuse (4+) BRAF cytoplasmic staining. C, Metastatic papillary thyroid microcarcinoma in a lymph node. D, Diffuse BRAF cytoplasmic staining in the metastatic thyroid carcinoma (hematoxylin-eosin, original magnification X200 [A and C]; original magnification X400 [B and D]).
Table 1. Summary of Hector Battifora Mesothelia-1 (HBME-1) Studies
(a, d)

                                             PTC, No.
Source, y                                      (%)

Barroeta et al, (114) 2006                  10/11 (91)
Park et al, (102) 2007                     166/181 (92)
Scognamiglio et al, (112) 2006              43/49 (88)
Nasr et al, (115) 2006                      49/51 (96)
Prasad et al, (108) 2005                    57/67 (85)
Cheung et al, (118) 2001                   76/138 (55)
Liu et al, (109) 2008                       39/53 (74)
Rossi et al, (107) 2006                     39/42 (93)
Saggiorato et al, (106) 2005                39/42 (93)
Rossi et al, (120) 2013
de Matos et al, (110) 2005                  79/84 (94)
Nechifor-Boila et al, (121) 2013            94/98 (96)
Torregrossa et al, (122) 2007              190/200 (95)
Volante et al, (104) 2004                  28/32 (88)e
Mai et al, (116) 2002 (f)                  42/42 (100)
Mase et al, (123) 2003                      35/36 (97)
Papotti et al, (124) 2005
Ito et al, (125) 2005                      37/37 (100)
Saleh et al, (95) 2010                      18/20 (90)

                                             1/3 (33)
                                            17/35 (49)
                                             2/49 (4)
                                             0/6 (0)
                                            2/21 (10)
                                             0/35 (0)
                                             1/12 (9)
                                             1/17 (6)
                                             2/50 (4)

0/5 (0)                                     10/18 (56)
                                            4/13 (31)

                                            19/45 (42)
4/4 (100)                                   17/62 (27)
                                             1/15 (7)
                                           47/155 (30)
                                            26/46 (57)

                                              FVPTC,
Source, y                                    No. (%)

Barroeta et al, (114) 2006                   3/4 (75)
Park et al, (102) 2007
Scognamiglio et al, (112) 2006              25/29 (86)
Nasr et al, (115) 2006                      9/10 (90)
Prasad et al, (108) 2005
Cheung et al, (118) 2001                    38/84 (45)
Liu et al, (109) 2008                       8/11 (73)
Rossi et al, (107) 2006                     12/14 (86)
Saggiorato et al, (106) 2005
Rossi et al, (120) 2013
de Matos et al, (110) 2005                  21/25 (84)
Nechifor-Boila et al, (121) 2013            73/90 (81)
Torregrossa et al, (122) 2007              128/138 (93)
Volante et al, (104) 2004
Mai et al, (116) 2002 (f)
Mase et al, (123) 2003
Papotti et al, (124) 2005                  14/14 (100)
Ito et al, (125) 2005
Saleh et al, (95) 2010                      11/12 (92)

0/5 (0)                                     9/10 (90)

                                            6/50 (12)
                                             4/48 (8)
4/4 (100)

                                              FTC,
Source, y                                    No. (%)

Barroeta et al, (114) 2006                  5/7 (71)
Park et al, (102) 2007                     22/25 (88)
Scognamiglio et al, (112) 2006
Nasr et al, (115) 2006
Prasad et al, (108) 2005                    3/6 (50)
Cheung et al, (118) 2001                    2/4 (50)
Liu et al, (109) 2008                       2/13 (15)
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005               21/33 (64)
Rossi et al, (120) 2013
de Matos et al, (110) 2005                 24/38 (63)
Nechifor-Boila et al, (121) 2013            4/9 (44)
Torregrossa et al, (122) 2007
Volante et al, (104) 2004
Mai et al, (116) 2002 (f)                  19/29 (66)
Mase et al, (123) 2003                     33/39 (85)
Papotti et al, (124) 2005
Ito et al, (125) 2005                      84/138 (61)
Saleh et al, (95) 2010                     18/22 (82)

                                            1/18 (6)
                                           11/54 (20)

                                            0/14 (0)
                                            1/29 (3)
                                            0/40 (0)
                                            0/14 (0)
                                            0/41 (0)

0/5 (0)                                     4/12 (33)

4/4 (100)                                   8/62 (13)

                                           17/98 (17)
                                            9/52 (17)

                                              PDC,
Source, y                                    No. (%)

Barroeta et al, (114) 2006                  3/3 (100)
Park et al, (102) 2007
Scognamiglio et al, (112) 2006
Nasr et al, (115) 2006
Prasad et al, (108) 2005
Cheung et al, (118) 2001                    4/6 (67)
Liu et al, (109) 2008
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005
Rossi et al, (120) 2013                    15/15 (100)
de Matos et al, (110) 2005
Nechifor-Boila et al, (121) 2013
Torregrossa et al, (122) 2007
Volante et al, (104) 2004
Mai et al, (116) 2002 (f)
Mase et al, (123) 2003
Papotti et al, (124) 2005
Ito et al, (125) 2005
Saleh et al, (95) 2010

                                            2/12 (17)

                                            4/19 (21)

0/5 (0)

4/4 (100)

                                             UDC,
Source, y                                   No. (%)

Barroeta et al, (114) 2006                 4/5 (80)
Park et al, (102) 2007
Scognamiglio et al, (112) 2006
Nasr et al, (115) 2006
Prasad et al, (108) 2005                    0/4 (0)
Cheung et al, (118) 2001                   1/2 (50)
Liu et al, (109) 2008
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005
Rossi et al, (120) 2013                     0/9 (0)
de Matos et al, (110) 2005                  0/2 (0)
Nechifor-Boila et al, (121) 2013
Torregrossa et al, (122) 2007
Volante et al, (104) 2004
Mai et al, (116) 2002 (f)
Mase et al, (123) 2003                     2/2 (100)
Papotti et al, (124) 2005
Ito et al, (125) 2005
Saleh et al, (95) 2010

                                            0/8 (0)
                                           0/14 (0)

                                           0/10 (0)

0/5 (0)

4/4 (100)

                                              HCC,
Source, y                                   No. (%)

Barroeta et al, (114) 2006                  1/4 (25)
Park et al, (102) 2007
Scognamiglio et al, (112) 2006
Nasr et al, (115) 2006
Prasad et al, (108) 2005                    1/8 (13)
Cheung et al, (118) 2001                    2/7 (29)
Liu et al, (109) 2008
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005
Rossi et al, (120) 2013
de Matos et al, (110) 2005
Nechifor-Boila et al, (121) 2013
Torregrossa et al, (122) 2007
Volante et al, (104) 2004                  26/70 (37)
Mai et al, (116) 2002 (f)                  2/12 (17)
Mase et al, (123) 2003
Papotti et al, (124) 2005
Ito et al, (125) 2005
Saleh et al, (95) 2010

                                            1/4 (25)

                                            0/10 (0)
                                            0/59 (0)

0/5 (0)                                    0/170 (0)

4/4 (100)

                                              Neg

Abbreviations: FA, follicular adenoma;FTC, follicular thyroid
carcinoma; FVPTC, follicular variant papillary thyroid
carcinoma; Graves, Graves disease;HA, Hurthle cell adenoma;HCC,
Hurthle cell carcinoma;LTis, lymphocytic thyroiditis;MC, medullary
thyroid carcinoma;Neg, negative; NG, nodular goiter;NL, normal
thyroid tissue;PDC, poorly differentiated thyroid carcinoma;PTC,
papillary thyroid carcinoma;UDC, undifferentiated thyroid carcinoma
(anaplastic carcinoma).

(a) Asa, (128) 2005: HBME-1 was expressed in 40% of malignant thyroid
tumors, including PTC and FTC.

(b) Zhu et al, (126) 2010: HBME-1 expression was identified in 95%
(148 of 155) of PTCs.

(c) Barut et al, (105) 2010: HBME-1 expression was identified in 72%
(47 of 65) of malignant thyroid neoplasms;rare focal reactivity was
identified in benign lesions.

(d) Wiseman et al, (127) 2008: HBME-1 expression was identified in
54% (53 of 99) of malignant thyroid neoplasms;only 5% (5 of 100) of
benign lesions.

(e) Hurthle cell variant PTC.

(f) Mai et al, (116) 2002: In addition, HBME-1 expression in Hurthle
cell PTC was 38% (5 of 13).

Table 1. Summary of Hector Battifora Mesothelia-1 (HBME-1)
Studies (a, d)

                                             PTC, No.
Source, y                                      (%)

Barroeta et al, (114) 2006                  10/11 (91)
Park et al, (102) 2007                     166/181 (92)
Scognamiglio et al, (112) 2006              43/49 (88)
Nasr et al, (115) 2006                      49/51 (96)
Prasad et al, (108) 2005                    57/67 (85)
Cheung et al, (118) 2001                   76/138 (55)
Liu et al, (109) 2008                       39/53 (74)
Rossi et al, (107) 2006                     39/42 (93)
Saggiorato et al, (106) 2005                39/42 (93)
Rossi et al, (120) 2013
de Matos et al, (110) 2005                  79/84 (94)
Nechifor-Boila et al, (121) 2013            94/98 (96)
Torregrossa et al, (122) 2007              190/200 (95)
Volante et al, (104) 2004                  28/32 (88)e
Mai et al, (116) 2002 (f)                  42/42 (100)
Mase et al, (123) 2003                      35/36 (97)
Papotti et al, (124) 2005
Ito et al, (125) 2005                      37/37 (100)
Saleh et al, (95) 2010                      18/20 (90)

                                             1/3 (33)
                                            17/35 (49)
                                             2/49 (4)
                                             0/6 (0)
                                            2/21 (10)
                                             0/35 (0)
                                             1/12 (9)
                                             1/17 (6)
                                             2/50 (4)

0/5 (0)                                     10/18 (56)
                                            4/13 (31)

                                            19/45 (42)
4/4 (100)                                   17/62 (27)
                                             1/15 (7)
                                           47/155 (30)
                                            26/46 (57)

                                              FVPTC,
Source, y                                    No. (%)

Barroeta et al, (114) 2006                   3/4 (75)
Park et al, (102) 2007
Scognamiglio et al, (112) 2006              25/29 (86)
Nasr et al, (115) 2006                      9/10 (90)
Prasad et al, (108) 2005
Cheung et al, (118) 2001                    38/84 (45)
Liu et al, (109) 2008                       8/11 (73)
Rossi et al, (107) 2006                     12/14 (86)
Saggiorato et al, (106) 2005
Rossi et al, (120) 2013
de Matos et al, (110) 2005                  21/25 (84)
Nechifor-Boila et al, (121) 2013            73/90 (81)
Torregrossa et al, (122) 2007              128/138 (93)
Volante et al, (104) 2004
Mai et al, (116) 2002 (f)
Mase et al, (123) 2003
Papotti et al, (124) 2005                  14/14 (100)
Ito et al, (125) 2005
Saleh et al, (95) 2010                      11/12 (92)

0/5 (0)                                     9/10 (90)

                                            6/50 (12)
                                             4/48 (8)
4/4 (100)

                                              FTC,
Source, y                                    No. (%)

Barroeta et al, (114) 2006                  5/7 (71)
Park et al, (102) 2007                     22/25 (88)
Scognamiglio et al, (112) 2006
Nasr et al, (115) 2006
Prasad et al, (108) 2005                    3/6 (50)
Cheung et al, (118) 2001                    2/4 (50)
Liu et al, (109) 2008                       2/13 (15)
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005               21/33 (64)
Rossi et al, (120) 2013
de Matos et al, (110) 2005                 24/38 (63)
Nechifor-Boila et al, (121) 2013            4/9 (44)
Torregrossa et al, (122) 2007
Volante et al, (104) 2004
Mai et al, (116) 2002 (f)                  19/29 (66)
Mase et al, (123) 2003                     33/39 (85)
Papotti et al, (124) 2005
Ito et al, (125) 2005                      84/138 (61)
Saleh et al, (95) 2010                     18/22 (82)

                                            1/18 (6)
                                           11/54 (20)

                                            0/14 (0)
                                            1/29 (3)
                                            0/40 (0)
                                            0/14 (0)
                                            0/41 (0)

0/5 (0)                                     4/12 (33)

4/4 (100)                                   8/62 (13)

                                           17/98 (17)
                                            9/52 (17)

                                              PDC,
Source, y                                    No. (%)

Barroeta et al, (114) 2006                  3/3 (100)
Park et al, (102) 2007
Scognamiglio et al, (112) 2006
Nasr et al, (115) 2006
Prasad et al, (108) 2005
Cheung et al, (118) 2001                    4/6 (67)
Liu et al, (109) 2008
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005
Rossi et al, (120) 2013                    15/15 (100)
de Matos et al, (110) 2005
Nechifor-Boila et al, (121) 2013
Torregrossa et al, (122) 2007
Volante et al, (104) 2004
Mai et al, (116) 2002 (f)
Mase et al, (123) 2003
Papotti et al, (124) 2005
Ito et al, (125) 2005
Saleh et al, (95) 2010

                                            2/12 (17)

                                            4/19 (21)

0/5 (0)

4/4 (100)

                                             UDC,
Source, y                                   No. (%)

Barroeta et al, (114) 2006                 4/5 (80)
Park et al, (102) 2007
Scognamiglio et al, (112) 2006
Nasr et al, (115) 2006
Prasad et al, (108) 2005                    0/4 (0)
Cheung et al, (118) 2001                   1/2 (50)
Liu et al, (109) 2008
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005
Rossi et al, (120) 2013                     0/9 (0)
de Matos et al, (110) 2005                  0/2 (0)
Nechifor-Boila et al, (121) 2013
Torregrossa et al, (122) 2007
Volante et al, (104) 2004
Mai et al, (116) 2002 (f)
Mase et al, (123) 2003                     2/2 (100)
Papotti et al, (124) 2005
Ito et al, (125) 2005
Saleh et al, (95) 2010

                                            0/8 (0)
                                           0/14 (0)

                                           0/10 (0)

0/5 (0)

4/4 (100)

                                              HCC,
Source, y                                   No. (%)

Barroeta et al, (114) 2006                  1/4 (25)
Park et al, (102) 2007
Scognamiglio et al, (112) 2006
Nasr et al, (115) 2006
Prasad et al, (108) 2005                    1/8 (13)
Cheung et al, (118) 2001                    2/7 (29)
Liu et al, (109) 2008
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005
Rossi et al, (120) 2013
de Matos et al, (110) 2005
Nechifor-Boila et al, (121) 2013
Torregrossa et al, (122) 2007
Volante et al, (104) 2004                  26/70 (37)
Mai et al, (116) 2002 (f)                  2/12 (17)
Mase et al, (123) 2003
Papotti et al, (124) 2005
Ito et al, (125) 2005
Saleh et al, (95) 2010

                                            1/4 (25)

                                            0/10 (0)
                                            0/59 (0)

0/5 (0)                                    0/170 (0)

4/4 (100)

                                              Neg

Abbreviations: FA, follicular adenoma;FTC, follicular thyroid
carcinoma; FVPTC, follicular variant papillary thyroid
carcinoma; Graves, Graves disease;HA, Hurthle cell adenoma;HCC,
Hurthle cell carcinoma;LTis, lymphocytic thyroiditis;MC, medullary
thyroid carcinoma;Neg, negative; NG, nodular goiter;NL, normal
thyroid tissue;PDC, poorly differentiated thyroid carcinoma;PTC,
papillary thyroid carcinoma;UDC, undifferentiated thyroid carcinoma
(anaplastic carcinoma).

(a) Asa, (128) 2005: HBME-1 was expressed in 40% of malignant thyroid
tumors, including PTC and FTC.

(b) Zhu et al, (126) 2010: HBME-1 expression was identified in 95%
(148 of 155) of PTCs.

(c) Barut et al, (105) 2010: HBME-1 expression was identified in 72%
(47 of 65) of malignant thyroid neoplasms;rare focal reactivity was
identified in benign lesions.

(d) Wiseman et al, (127) 2008: HBME-1 expression was identified in
54% (53 of 99) of malignant thyroid neoplasms;only 5% (5 of 100) of
benign lesions.

(e) Hurthle cell variant PTC.

(f) Mai et al, (116) 2002: In addition, HBME-1 expression in
Hurthle cell PTC was 38% (5 of 13).

Table 2. Summary of Cytokeratin 19 (CK19) Studies (a-b)

                                       PTC,          FVPTC,
Source, y                             No. (%)        No. (%)

Barroeta et al, (114) 2006          10/11 (91)      4/4 (100)
Park et al, (102) 2007             175/181 (97)
Scognamiglio et al, (112) 2006      49/49 (100)    26/29 (90)
Nasr et al, (115) 2006              51/51 (100)    10/10 (100)
Prasad et al, (108) 2005            48/67 (72)
Erkilic et al, (131) 2002           25/25 (100)
Sahoo et al, (132) 2001             15/15 (100)    10/10 (100)
Cheung et al, (118) 2001            91/138 (66)    48/84 (57)
Liu et al, (109) 2008               41/53 (78)      2/11 (22)
Murphy et al, (130) 2008            20/20 (100)     2/9 (18)
Rossi et al, (107) 2006             36/42 (86)     11/14 (79)
Saggiorato et al, (106) 2005        37/42 (88)
Beesley and McLaren, (111) 2002     26/26 (100)
Song et al, (133) 2011             425/441 (96)
de Matos et al, (110) 2005          61/84 (73)     13/25 (52)
Baloch et al, (129) 1999            39/39 (100)    26/26 (100)
Bose et al, (134) 2012              22/22 (100)
Saleh et al, (95) 2010              17/20 (85)     10/12 (83)
                                      0/3 (0)       0/3 (0%)
                                    10/35 (29)
                                     7/49 (14)
                                     5/6 (83)
                                     1/21 (5)

                                    20/20 (100)
                                     7/35 (20)
                                     0/12 (0)
                                     4/15 (27)      0/11 (0%)
                                     1/17 (6)
                                     5/50 (10)
2/2 (100%)                           5/20 (25)
                                   39/151 (26%)i
1/5 (20)                             6/18 (33)      6/10 (60)
                                      0/4 (0)
                                     6/8 (75)
                                    23/46 (50)

                                      FTC,         PDC,        UDC,
Source, y                           No. (%)      No. (%)      No. (%)

Barroeta et al, (114) 2006          3/7 (43)     2/3 (67)    4/5 (80)
Park et al, (102) 2007             11/25 (44)
Scognamiglio et al, (112) 2006
Nasr et al, (115) 2006
Prasad et al, (108) 2005            3/6 (50)                 1/4 (25)
Erkilic et al, (131) 2002
Sahoo et al, (132) 2001
Cheung et al, (118) 2001            0/4 (0)      3/6 (50)     0/2 (0)
Liu et al, (109) 2008               0/13 (0)
Murphy et al, (130) 2008           6/14 (43)
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005       23/33 (70)
Beesley and McLaren, (111) 2002    5/12 (42)                 1/1 (100)
Song et al, (133) 2011
de Matos et al, (110) 2005         8/38 (21)                  0/2 (0)
Baloch et al, (129) 1999            0/2 (0)
Bose et al, (134) 2012
Saleh et al, (95) 2010             19/22 (86)
                                   3/18 (17)    8/12 (67)    6/13 (46)
                                    5/54 (9)

                                   6/14 (43)    16/19 (84)   2/8 (25)

                                   5/25 (20)

                                   8/40 (20)
                                    0/14 (0)                 1/10 (10)

                                    1/41 (2)

2/2 (100%)                          2/8 (25)

1/5 (20)                           2/12 (17)
                                    0/5 (0)
                                    4/8 (50)
                                   8/52 (15)

                                      HCC,
Source, y                            No. (%)

Barroeta et al, (114) 2006          1/4 (25)
Park et al, (102) 2007
Scognamiglio et al, (112) 2006
Nasr et al, (115) 2006
Prasad et al, (108) 2005            4/8 (50)
Erkilic et al, (131) 2002
Sahoo et al, (132) 2001
Cheung et al, (118) 2001            2/7 (29)
Liu et al, (109) 2008
Murphy et al, (130) 2008
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005
Beesley and McLaren, (111) 2002
Song et al, (133) 2011
de Matos et al, (110) 2005
Baloch et al, (129) 1999
Bose et al, (134) 2012
Saleh et al, (95) 2010
                                    1/4 (25)

                                   10/10 (100)

                                    8/19 (42)

2/2 (100%)

1/5 (20)                            0/170 (0)
                                       Neg

Abbreviations: FA, follicular adenoma; FTC, follicular thyroid
carcinoma; FVPTC, follicular variant papillary thyroid carcinoma;
Graves, Graves disease; HA, Hurthle cell adenoma; HCC, Hurthle cell
carcinoma; LTis, lymphocytic thyroiditis; MC, medullary thyroid
carcinoma; Neg, negative;  NG, nodular goiter; NL, normal thyroid
tissue; PDC, poorly differentiated thyroid carcinoma; PTC, papillary
thyroid carcinoma; UDC, undifferentiated thyroid carcinoma
(anaplastic carcinoma).

(a) Asa, (128) 2005: CK19 expression identified in 60% of PTC cases;
nonreactive or focal positivity in follicular neoplasm and NG,
usually inflamed tissue.

(b) Zhu et al, (126) 2010: CK19 expression was identified in 87% (135
of 155) of PTC cases.

(c) Nechifor-Boila et al, (121) 2013: CK19 expressed in 46% (93 of
204) of PTC cases.

(d) Cameron and Berean, (234) 2003: Diffuse CK19 expression is
identified in 100% of PTCs, including FVPTC; FTC, FA, and NG were
negative or showed focal reactivity.

(e) Casey et al, (135) 2003: CK19 expression is identified in 100%
(30 of 30) of malignant tumors, however with significant reactivity
in nonneoplastic thyroid tissue.

(f) Barut et al, (105) 2010: CK19 expression identified in 77% (50 of
65) of malignant thyroid tumors; only focal reactivity noted in
benign lesions.

(g) Wiseman et al, (127) 2008: CK19 is expressed in 97% (96 of 99)
of malignant tumors.

(h) Nakamura et al, (100) 2006: A panel consisting of Hector
Battifora mesothelial/1 (HBME/1), galectin/3 (GAL/3), and CK19 or
HBME/1, Cbp/ p300- interacting transactivator with Glu-Asp-rich
carboxy-terminal domain, 1 (CITED-1), and GAL-3: useful in
distinguishing FA from FVPTC.

(i) One hundred fifty-one cases of NG and FA.

Table 3. Summary of Galectin-3 (GAL-3) Studies (a-e)

                                      PTC,         FVPTC,
Source, y                           No. (%)        No. (%)

Barroeta et al, (114) 2006          9/11 (82)      3/4 (75)
Park et al, (102) 2007            179/181 (99)
Scognamiglio et al, (112) 2006     47/49 (96)    26/29 (90)
Prasad et al, (108) 2005           63/67 (94)
Liu et al, (109) 2008              49/53 (92)     4/11 (36)
Murphy et al, (130) 2008          20/20 (100)      5/9 (55)
Rossi et al, (107) 2006            37/42 (88)    10/14 (71)
Saggiorato et al, (106) 2005      42/42 (100)
Beesley and McLaren, (111) 2002    22/26 (85)
Song et al, (133) 2011            427/441 (97)
Rossi et al, (120) 2013
de Matos et al, (110) 2005         61/84 (73)    13/25 (52)
Weber et al, (138) 2004            22/24 (92)
Saggiorato et al, (139) 2004      26/26 (100)
Orlandi et al, (140) 1998         18/18 (100)
Saggiorato et al, (141) 2001
Savin et al, (103) 2008           126/147 (86)
Bartolazzi et al, (142) 2008                     67/86 (78)
Torregrossa et al, (122) 2007     152/200 (76)   90/138 (65)
Volante et al, (104) 2004         31/32 (97)g
Volante et al, (117) 2004
Gaffney et al, (143) 2003          55/60 (92)
Coli et al, (144) 2002            28/28 (100)    10/10 (100)
Nascimento et al, (145) 2001        9/11 (82)
Papotti et al, (124) 2005                        13/14 (93)
Saleh et al, (95) 2010             18/20 (90)    10/12 (83)
Bartolazzi et al, (146) 2001      195/201 (97)

                                     FTC,          PDC,
Source, y                           No. (%)       No. (%)

Barroeta et al, (114) 2006          4/7 (57)      1/3 (33)
Park et al, (102) 2007            16/25 (64)
Scognamiglio et al, (112) 2006
Prasad et al, (108) 2005            4/6 (67)
Liu et al, (109) 2008              4/13 (31)
Murphy et al, (130) 2008           3/14 (21)
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005      29/33 (88)
Beesley and McLaren, (111) 2002   12/12 (100)
Song et al, (133) 2011
Rossi et al, (120) 2013                         15/15 (100)
de Matos et al, (110) 2005         8/38 (21)
Weber et al, (138) 2004             4/9 (44)
Saggiorato et al, (139) 2004      39/39 (100)
Orlandi et al, (140) 1998         17/17 (100)
Saggiorato et al, (141) 2001      17/17 (100)
Savin et al, (103) 2008           10/18 (56)
Bartolazzi et al, (142) 2008      11/15 (73)      5/7 (71)
Torregrossa et al, (122) 2007
Volante et al, (104) 2004
Volante et al, (117) 2004
Gaffney et al, (143) 2003         14/21 (67)
Coli et al, (144) 2002              3/5 (60)
Nascimento et al, (145) 2001      11/14 (79)
Papotti et al, (124) 2005
Saleh et al, (95) 2010            18/22 (82)
Bartolazzi et al, (146) 2001      54/57 (95)    13/20 (65)

                                     UDC,         HCC,         MC,
Source, y                          No. (%)       No. (%)     No. (%)

Barroeta et al, (114) 2006         4/5 (80)      3/4 (75)
Park et al, (102) 2007
Scognamiglio et al, (112) 2006
Prasad et al, (108) 2005          4/4 (100)      7/8 (88)
Liu et al, (109) 2008
Murphy et al, (130) 2008
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005
Beesley and McLaren, (111) 2002     0/1 (0)                  1/2 (50)
Song et al, (133) 2011
Rossi et al, (120) 2013             0/9 (0)
de Matos et al, (110) 2005          0/2 (0)                  0/5 (0)
Weber et al, (138) 2004
Saggiorato et al, (139) 2004
Orlandi et al, (140) 1998
Saggiorato et al, (141) 2001
Savin et al, (103) 2008
Bartolazzi et al, (142) 2008                   18/22 (82)
Torregrossa et al, (122) 2007
Volante et al, (104) 2004                      66/70 (94)
Volante et al, (117) 2004                      46/49 (94)
Gaffney et al, (143) 2003
Coli et al, (144) 2002            1/1 (100)     1/1 (100)
Nascimento et al, (145) 2001                   10/17 (59)
Papotti et al, (124) 2005
Saleh et al, (95) 2010
Bartolazzi et al, (146) 2001      18/20 (90)   13/13 (100)   3/7 (43)

                                      FA,           HA,
Source, y                           No. (%)       No. (%)

Barroeta et al, (114) 2006                       1/3 (33)
Park et al, (102) 2007               1/35 (3)
Scognamiglio et al, (112) 2006      9/49 (18)
Prasad et al, (108) 2005            2/21 (10)
Liu et al, (109) 2008                0/12 (0)
Murphy et al, (130) 2008            5/15 (33)    7/11 (64)
Rossi et al, (107) 2006              0/17 (0)
Saggiorato et al, (106) 2005         3/50 (6)
Beesley and McLaren, (111) 2002     2/20 (10)
Song et al, (133) 2011            77/151 (51)f
Rossi et al, (120) 2013
de Matos et al, (110) 2005           2/18(11)    7/10 (70)
Weber et al, (138) 2004             4/13 (31)
Saggiorato et al, (139) 2004        2/105 (2)
Orlandi et al, (140) 1998           3/29 (10)
Saggiorato et al, (141) 2001         4/52 (8)
Savin et al, (103) 2008             4/28 (14)
Bartolazzi et al, (142) 2008       19/176(11)
Torregrossa et al, (122) 2007
Volante et al, (104) 2004                        6/50 (12)
Volante et al, (117) 2004                        6/45 (13)
Gaffney et al, (143) 2003           3/14 (21)
Coli et al, (144) 2002             17/27 (63)
Nascimento et al, (145) 2001         1/9 (11)    1/14 (7)
Papotti et al, (124) 2005            3/15 (2)
Saleh et al, (95) 2010             19/46 (41)
Bartolazzi et al, (146) 2001        4/125 (3)    1/7 (14)

                                     NG,         LTis,       Graves,
Source, y                          No. (%)      No. (%)      No. (%)

Barroeta et al, (114) 2006         1/18 (6)                 6/13 (46)
Park et al, (102) 2007             3/54 (6)
Scognamiglio et al, (112) 2006
Prasad et al, (108) 2005          16/29 (55)                1/14 (7)
Liu et al, (109) 2008              0/14 (0)                 1/10 (10)
Murphy et al, (130) 2008
Rossi et al, (107) 2006            0/41 (0)
Saggiorato et al, (106) 2005
Beesley and McLaren, (111) 2002    3/8 (38)
Song et al, (133) 2011
Rossi et al, (120) 2013
de Matos et al, (110) 2005         1/12 (8)
Weber et al, (138) 2004
Saggiorato et al, (139) 2004
Orlandi et al, (140) 1998
Saggiorato et al, (141) 2001
Savin et al, (103) 2008
Bartolazzi et al, (142) 2008      3/126 (2)
Torregrossa et al, (122) 2007
Volante et al, (104) 2004
Volante et al, (117) 2004          0/13 (0)    14/15 (93)
Gaffney et al, (143) 2003
Coli et al, (144) 2002            7/25 (28)
Nascimento et al, (145) 2001       0/30 (0)     0/11 (0)
Papotti et al, (124) 2005
Saleh et al, (95) 2010            8/52 (15)
Bartolazzi et al, (146) 2001       0/50 (0)     2/29 (7)     0/1 (0)

                                     NL,
Source, y                          No. (%)

Barroeta et al, (114) 2006
Park et al, (102) 2007
Scognamiglio et al, (112) 2006
Prasad et al, (108) 2005          0/59 (0)
Liu et al, (109) 2008
Murphy et al, (130) 2008          8/19 (42)
Rossi et al, (107) 2006
Saggiorato et al, (106) 2005
Beesley and McLaren, (111) 2002
Song et al, (133) 2011
Rossi et al, (120) 2013
de Matos et al, (110) 2005        0/170 (0)
Weber et al, (138) 2004
Saggiorato et al, (139) 2004
Orlandi et al, (140) 1998
Saggiorato et al, (141) 2001
Savin et al, (103) 2008           0/142 (0)
Bartolazzi et al, (142) 2008
Torregrossa et al, (122) 2007
Volante et al, (104) 2004
Volante et al, (117) 2004
Gaffney et al, (143) 2003
Coli et al, (144) 2002
Nascimento et al, (145) 2001      0/18 (0)
Papotti et al, (124) 2005
Saleh et al, (95) 2010
Bartolazzi et al, (146) 2001      0/75 (0)

Abbreviations: FA, follicular adenoma; FTC, follicular thyroid
carcinoma; FVPTC, follicular variant papillary thyroid carcinoma;
Graves, Graves disease; HA, Hurthle cell adenoma; HCC, Hurthle cell
carcinoma; LTis, lymphocytic thyroiditis; MC, medullary thyroid
carcinoma; NG, nodular goiter; NL, normal thyroid tissue; PDC,
poorly differentiated thyroid carcinoma; PTC, papillary thyroid
carcinoma; UDC, undifferentiated thyroid carcinoma (anaplastic
carcinoma).

(a) Zhu et al, (126) 2010: GAL-3 expressed in 92% (142 of 155) of
PTCs.

(b) Barut et al, (105) 2010: GAL-3 expressed in 72% (47 of 65) of
malignant thyroid tumors.

(c) Wiseman et al, (127) 2008: GAL-3 expressed in 90% (89 of 99) of
malignant thyroid tumors; 29% (29 of 100) of benign thyroid
lesions.

(d) Asa, (128) 2005: CK19 is a marker of malignancy; inflamed NL
and NG also showed reactivity.

(e) Saggiorato et al, (139) 2004; 0rlandi et al, (140) 1998; and
Saggiorato et al, (141) 2001: Studies were conducted by using
fine-needle aspiration samples, with follow-up histologic tissue
confirmation. Data provided in this table are study results on
tissue.

(f) Included NG and FA.

(g) Hurthle cell variant PTC.

Table 4. Trophoblastic Cell Surface Antigen (TROP2) Expression
in 136 Cases of Thyroid Neoplasms

Diagnosis      1 +   2+   3+   4+   Total Positive Cases (%)

PTC (n = 48)    5    10   18   10           43 (90)
FA (n = 51)     0    0    0    0             0 (0)
FTC (n = 37)    0    0    0    0             0 (0)

Abbreviations: FA, follicular adenoma; FTC, follicular thyroid
carcinoma; PTC, papillary thyroid carcinoma.

Table 5. Trophoblastic Cell Surface Antigen (TROP2) Expression
in 61 Cases of Atypical Follicular Lesions

Diagnosis (n)   TROP2, No. (%)   CK19, No. (%)

PTC (33)           23 (70)          22 (67)
AFN (11)            0 (0)           2 (18)
ANFNA (17)          0 (0)           3 (18)

Diagnosis (n)   HBME-1, No. (%)   GAL-3, No. (%)

PTC (33)           33 (100)          33 (100)
AFN (11)            9 (82)            8 (73)
ANFNA (17)          7 (41)            6 (35)

Abbreviations: AFN, atypical follicular neoplasm; ANFNA, adenomatoid
nodule with focal nuclear atypia; CK19, cytokeratin 19; GAL-3,
galectin-3;  HBME-1, Hector Battifora mesothelial-1; n, number of
cases; PTC, papillary thyroid carcinoma.

Table 6. Summary of Studies With Thyroperoxidase (TPO),
Monoclonal Antibody 47

                                 Cutoff, (a)        PTC,
Source, y                             %           No. (%)

Weber et al, (138) 2004               5        12/24 (50) (b)
Savin et al, (103) 2008              50          13/147 (9)

Yousaf et al, (165) 2008             80           0/17 (0)

Christensen et al, (166) 2000        80         0/27 (0) (e)

De Micco et al, (167) 1994 (a)       80

De Micco et al, (164) 1991 (g)       80           4/43 (9)

de Micco et al, (168) 2008 (i)       80           0/59 (0)

Savin et al, (169) 2006              80

                                                UDC,   MC,
                                     FTC,       No.    No.
Source, y                          No. (%)      (%)    (%)

Weber et al, (138) 2004          8/9 (89) (c)
Savin et al, (103) 2008           10/18 (56)

Yousaf et al, (165) 2008         3/6 (50) (d)   0/3    0/1
                                                (0)    (0)
Christensen et al, (166) 2000

De Micco et al, (167) 1994 (a)     0/9 (0)

De Micco et al, (164) 1991 (g)    7/22 (32)

de Micco et al, (168) 2008 (i)     0/32 (0)

Savin et al, (169) 2006           11/52 (21)

                                                HA,     NG,
                                     FA,        No.     No.
Source, y                          No. (%)      (%)     (%)

Weber et al, (138) 2004          13/13 (100)
Savin et al, (103) 2008          23/28 (82)

Yousaf et al, (165) 2008         30/31 (97)            67/67
                                                       (100)
Christensen et al, (166) 2000    25/26 (96)

De Micco et al, (167) 1994 (a)   50/60 (83)    8/17
                                               (47)
De Micco et al, (164) 1991 (g)   49/50 (98)    10/10   10/10
                                               (100)   (100)
de Micco et al, (168) 2008 (i)   39/54 (72)            51/55
                                                       (93)
Savin et al, (169) 2006          24/40 (60)

                                               NL,
                                  Graves,      No.
Source, y                         No. (%)      (%)

Weber et al, (138) 2004
Savin et al, (103) 2008                      94/114
                                              (83)
Yousaf et al, (165) 2008

Christensen et al, (166) 2000      71/71
                                 (100) (f)
De Micco et al, (167) 1994 (a)

De Micco et al, (164) 1991 (g)     10/10     100 (h)
                                   (100)
de Micco et al, (168) 2008 (i)

Savin et al, (169) 2006

Abbreviations: FA, follicular adenoma; FTC, follicular thyroid
carcinoma; Graves, Graves disease; HA, Hurthlecell adenoma; MC,
medullary thyroid carcinoma; NG, nodular goiter; NL, normal thyroid
tissue; PTC, papillary thyroid carcinoma; UDC, undifferentiated
thyroid carcinoma (anaplastic carcinoma).

(a) Cutoff: The percentage of cells showing immunoreactivity to
define positive.

(b) Seven of 12 TPO-positive cases showed 3+, diffuse staining; 5 of
12 were 3+ with fewer than 50% cells being positive. If 80% cutoff
used, which is used by all other studies, the positive rate would be
7 of 24 (29%).

(c) Seven of 8 TPO-positive cases showed 3+, diffuse staining;  1 of
8 was 1 + with fewer than 50% cells being positive. If 80% cutoff
used, the positive rate would be 7 of 9 (78%).

(d) The 3 TPO-positive cases are minimally invasive follicular
carcinomas with Hurthle cell morphology.

(e) Twenty-seven follicular malignant cases; not specified whether
PTC or follicular carcinoma.

(f) Seventy-one benign thyroid tissues, not specified.

(g) De Micco et al, (164) 1991: Fine-needle aspiration specimens
with confirmed diagnoses by surgical follow-up histology. Study was
performed on half of the slides.

(h) Number of cases was not specified.

(i) de Micco et al, (168) 2008: Study conducted on direct smears.

(j) The 40 cases of follicular adenomas include cases with Hurthle
cell morphology.

Table 7. Correlation of BRAF V600E Mutation in Papillary Thyroid
Carcinoma by Immunohistochemistry (IHC) and Molecular Testing

Source, y                    Antibody     IHC Result

Capper et al, (233) 2011     VE1          9 cases positive by IHC

Koperek et al, (232) 2012    VE1          39 cases positive by IHC

Bullock et al, (231) 2012    VE1          68 cases positive by IHC

Zagzag et al, (230) 2013     VE1          25 of 28 BRAF-mutated
                                          cases positive by IHC

McKelvie et al, (228) 2013   VE1          50 cases positive by IHC

Routhier et al, (227) 2013   VE1          13 cases positive by IHC

Routhier et al, (227) 2013   Anti-B-Raf   16 cases positive by IHC
                             mutation     including 3 cases that
                             mouse        were negative by
                             antibody     SNaPshot.

Source, y                    Molecular Testing     Comment

Capper et al, (233) 2011     9 cases positive by   100% sensitivity
                             PCR amplification     and specificity for
                             and direct            IHC; 3 additional
                             sequencing            IHC-positive cases;
                                                   molecular testing
                                                   failed.

Koperek et al, (232) 2012    39 cases positive     100% sensitivity
                             by gene sequencing    and specificity; 1
                                                   strong IHC-positive
                                                   cases was negative
                                                   by molecular
                                                   testing.

Bullock et al, (231) 2012    59 cases positive     In 11 discordant
                             for BRAF by Sanger    cases, further
                             sequencing            testing favored the
                                                   original IHC
                                                   results in 8 cases,
                                                   suggesting that IHC
                                                   was a more
                                                   sensitive method
                                                   than Sanger
                                                   sequencing.

Zagzag et al, (230) 2013     28 cases positive     Sensitivity 89% and
                             for BRAF by direct    specificity 100% by
                             sequencing            IHC.

McKelvie et al, (228) 2013   41 cases positive     8 of 9 IHC-positive
                             for BRAF by C-PCR     and C-PCR- negative
                                                   cases were positive
                                                   for BRAF by
                                                   SNaPshota method,
                                                   suggesting that IHC
                                                   was more sensitive
                                                   than C-PCR method.

Routhier et al, (227) 2013   13 cases positive     100% sensitivity
                             for BRAF by           and specificity by
                             SNaPshot genotyping   IHC.
                             assay

Routhier et al, (227) 2013   13 cases positive     100% sensitivity
                             for BRAF by           and 70% specificity
                             SNaPshot genotyping   by IHC.
                             assay

Abbreviations: BRAF, B-isoform of RAF kinase; C-PCR, competitive PCR;
PCR, polymerase chain reaction.
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Date:Jan 1, 2015
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