S100 can be used as a tumor marker in canine mammary tumors.
S100 protein, named from their solubility in a 100% saturated solution of ammonium sulfate at neutral pH, is one of the commonly used tumors markers used in immunohistochemistry. It is a multigenic family of nonubiquitous [Ca.sup.2+] - modulated proteins of the EF-hand type expressed in vertebrates exclusively and implicated in intracellular and extracellular regulatory activities. It has been well established that the genes for several S100 proteins are associated with cell differentiation, malignant transformation, and cell cycle growth, and that their expression levels are varied according to various growth- and growth-inhibitory conditions. S100 has been demonstrated by immunohistochemistry in healthy organs in animals such as in the spleens of bovine, sheep, and Philippine swamp buffalo, in the kidneys of sheep and goat, Philippine swamp buffalo, rat, goldfish, saltwater fishes, frog, lizards and ostrich, in testis of poultry and rabbits, Philippine swamp buffalo, and cat, epididymis of poultry and rabbits, and in ovary of rat. In addition, it was also immunohistochemically demonstrated in pathologic conditions in humans such as papillary carcinoma of thyroid, renal cell carcinoma, and nerve sheath tumors such as neurilemoma, neurofibromas, and myxoid sheath nerve tumor and in animals such as canine amelanotic melanoma. It has also been expressed in cases of acute and chronic inflammatory disorders, gastrointestinal inflammation, lung disorders, primary tumour such as melanoma, head-and-neck cancers, and breast cancer.
Canine mammary tumors (CMTs) are second to skin tumors as most frequent neoplasm in dogs, wherein approximately half of its cases are malignant, but prevalence is low in countries that routinely perform ovariectomy or ovariohysterectomy. In addition, these tumors were comparatively studied and said to have molecular and biological similarities with the human mammary tumors.[22-24] Based on this similarities, different classification methods of CMTs have been proposed in the literature, some of them are adapted from human classification systems.[25-28] Although diagnosis as well as prognosis in cases of mammary tumors remains a challenge, researches done on this subject had been of great help and had been an interest of research in the field of pathology and oncology.
Some of the diagnostic tools that are routinely used in tumors include physical examination, radiographic screening, and histopathological diagnosis through the routine use of hematoyxlin and eosin staining in surgical biopsy or aspiration biopsy, but there may be factors that may alter these morphologically-based diagnostics that may lead to confusion and to an erroneous diagnosis of malignancy. Thus, there have been suggestions on the use of immunohistochemistry which have been proven to be a valuable technique in research and diagnostic histopathology and cytology for the identification and classification of tumor cells using a wide variety of markers.[30,31]
As there are many different tumor markers, each are indicative of particular disease process, and they are used in oncology to help detect the presence of cancer. Particularly with S100 protein, diseases that are associated with altered expression levels of S100 can be classified into four categories as follows: Neurologic disorders, neoplastic disorders, cardiac diseases, and inflammatory diseases. S100 can be found in body fluids, including serum, urine, seminal plasma, saliva, sputum, cerebrospinal fluid and feces and abscess fluid, principally associated with active disease states. According to Donato et al.,  S100A8 or S100A9 complex and S100B are considered biomarkers for particular disease processes. An elevated level of a tumor marker can indicate cancer, but there can also be other causes of the elevation. Different forms of cancer exhibit dramatic changes in the expression of S100 proteins such as S100B, S100A2, S100A4, S100A6, and S100P. For example, elevated levels of S100A4 (metastasin) are associated with poor survival rates in human breast cancer patients and have been found to induce metastasis in mouse models. Another example is a high secretion of S100B in malignant melanoma, reflecting tumor load, stage, and prognosis. According to Sedaghat and Notopoulos, other members of the S100 protein family may prove to be useful biomarkers in future applications and may S100 protein-targeted therapies emerge as useful opportunities in specific clinical settings. Despite the emergence of newer markers reflecting differentiation, proliferation, immunomodulation, and other relevant processes, according to Ohsie et al., S100B remains to be the most sensitive immunohistochemical marker of melanoma. According to Schmitt and Bacchi, S-100 protein is also useful as a tumor marker in diagnostic immunocytochemistry. In their study, they detected S-100 protein by the immunoperoxidase technique in a heterogeneous group of 159 tumors to determine whether this marker may be of value in facilitating immunocytochemical diagnosis. The results have shown that S100 was widely distributed and demonstrated the strongest degrees of reactivity. S100 was identified in virtually all nerve sheath tumors such as neurilemoma, neurofibromas, myxoid sheath nerve tumor, and also in tumors of controversial histogenesis such as granular cell tumors. In addition, the great majority of carcinomas in their study did not express S100, with only two cases of breast carcinoma displaying focal S100 staining. Despite its presence in a wide array of cell types, it was concluded that S100 protein continues to be an extremely useful marker, especially for soft tissue and peripheral nervous system tumors.
By recognizing and understanding the signatures of normal cells and how these become cancerous, technology can provide important insights into the etiology of cancer that can be useful for early detection, diagnosis, and treatment. Therefore, biomarkers could be an invaluable tool for cancer detection, diagnosis, prognosis, and therapy selection. One of the advent laboratory diagnostic tools, immunohistochemistry, was used in this study, particularly in detecting S100 protein in mammary tumors. Since there had been challenges in diagnosing CMT, as a possible option to do immunohistochemistry in addition to the routine diagnostic method of hematoxylin and eosin staining for histopathology, this study could have a potential for further investigation for more precise diagnosis. When S100 protein will be proven to be a good tumor marker in the tumorous mammary organ in dogs, this study could be useful to veterinarians, practitioners, clinicians, and pathologists. This study could also be a reference for future researchers who are interested in developing more accurate diagnostic tools for various diseases such as tumors and further studying on other potential tumor markers. In addition, as there have been previous studies on S100 proteins in human specimens and in other animal species, this study could be of help in decreasing the limitation of its use in the veterinary field and may also serve as a diagnostic tool in the future and may be further studied for its use in classifying and prognosis of CMT.
MATERIALS AND METHODS
Ten mammary tumor specimens and a normal mammary tissue from dogs regardless of age and breed were studied. Ten mammary tumor specimens were obtained from bitches that underwent surgery at the different veterinary clinics and hospitals within Metro Manila, Philippines from December 2015 to March 2016. Consent from the owners as well as the Veterinarians were obtained before sampling. Normal mammary tissue were obtained from anatomical cadavers of De La Salle Araneta University. Immediately after surgical resection, tissue samples were collected from the middle portion of the tumor and fixed in 10% buffered formalin for 72 h. The same method was done for the normal lactating mammary gland sample. The fixed samples were then processed routinely using the paraffin technique, sectioned at 5 qm using an 820 rotary microtome (American Optical[R], New York) and mounted on ordinary slides for Hematoxylin-Eosin staining to identify various components of the mammary gland or on polysine-coated slides (Thermo Scientific, Massachusetts) for immunohistochemistry.
The avidin-biotin-peroxidase complex (ABC) method was used to detect immunoreactivity to S-100. Two tissue sections per sample were deparaffinized and washed with 0.1 M tris buffer saline (TBS) at pH 7.6, seven times at 5 min interval. The tissue sections were incubated with 20% normal goat serum (NGS) in TBS for 30 min under room temperature to decrease non-specific staining. Thereafter, the sections were incubated overnight with primary antibody, the polyclonal rabbit anti-S-100 (DakoCytomation, Copenhagen) at 1:2000 dilution with TBS containing 5% NGS at 4[degrees]C in a moisture chamber. The sections were washed with TBS four times at 5 min interval; incubated with biotinylated anti-rabbit IgG, 1:500 dilution (Vector Laboratories, California) in TBS containing 1.5% NGS, for 90 min at room temperature; washed in TBS three times at 5 min interval; incubated with ABC (Vector Laboratories, California) for 60 min at room temperature; and rinsed with TBS three times at 5 min interval. The S100 immunoreactivity was visualized through incubation of the tissue sections in a solution of imidazole-HCl buffer containing 0.05% 3,3'-diaminobenzidine, for 8 min at room temperature. Immunoreaction was stopped by washing the tissue sections in TBS 10 times until no further browning of the sections is occurring. The immunostained sections were placed in a covered plastic container to dry overnight at room temperature. Thereafter, the sections were rehydrated in triple distilled water twice, dehydrated in increasing concentrations of ethanol (70%, 90%, 100% and 100%) for 3 min per concentration, cleared in two changes of xylene for 3 min, and mounted with entellan (Eukitt[R]) (Merck KgaA, Darmstadt). The skin and skeletal muscle were used as a positive and negative control, with the same procedures performed for the canine mammary gland.
The tumors collected were classified according to the World Health Organization criteria for canine mammary lesions[25-28] based on the most pronounced histological pattern observed in more than 50% of the tumor mass. Whenever tumors displayed multiple morphological patterns, without more prominent growth pattern present in 50% of the tumor mass, lesions were classified as tumors with mixed morphology tumor. Tumor malignancy grade was determined using the Elston and Ellis scoring system based on the assessment of three morphological features: (1) The degree of glandular differentiation assessed using tubular formation, (2) nuclear pleomorphism, and (3) mitotic activity. Each parameter was graded into three categories to which a score of 1-3 was assigned. For evaluating the tubule formation, the tubular structures were scored qualitatively and quantitatively while the proportion occupied by such tubular structures was assessed semiquantitatively. One point is assigned when more than 75% of the area is composed of definite tubules. Two points are allocated for tumors in which between 10% and 75% of the area shows tubule formation. Where tubules occupy 10% or less of the tumor, three points are assigned. On the other hand, in assessing nuclear pleomorphism, one point is appropriate when the tumor nuclei are small regular uniform cells. Two points are given when the nuclei are larger than normal, have more open vesicular nuclei with visible, usually single, nucleoli, and there is moderate variation in size and shape. When there is marked variation in size and shape, especially when there are very large and bizarre nuclei present, three points is given. Finally, for the mitotic count, up to 7 mitosis per 10 high fields are scored 1 point, 8-16 are scored 2 points and more than 17 are scored 3 points. After the scores were added of each category, a number between 3 and 9 would be obtained then using the Elston grade, the following grade was allocated on the following basis: Grade I (well differentiated) is allocated for 3-5 points, Grade II (moderately differentiated) for 6-7 points and Grade III (poorly differentiated) for 8-9 points.
Tumor and normal lactating mammary gland sections were observed under a light microscope to identify S100 immunoreactive cells. The reactions were designated as positive (+) or negative (-). Photomicrographs of representative sections were taken using light microscope with digital camera (Olympus Camedia C-400) (Olympus UK Ltd., Hertfordshire) attachment for documentation.
Ten CMTs were evaluated using the hematoxylin and eosin and classified according to the histological classification of mammary tumors of the dog and cat [Table 1].
Three tumors were benign, six were malignant and one case of dysplasia. One normal mammary gland sample was used for comparative immunohistochemical evaluation. Immunohistochemical reactivity of the different cell types is summarized in Table 2.
Normal Mammary Gland
In the normal mammary gland of dog [Figure 1], the secretory parenchyma is well developed and the connective tissue is reduced. The lumens of the secretory glands and ducts are filled with secretion. The epithelial and myoepithelial cells were negatively immunostained by S100, and only the endothelial cells of vessels and blood were found to be immunoreactive. There was also a faint brown stain observed in the secretions within alveoli which are milk [Figure 2].
In the hematoxylin and eosin stain, it was described to have epithelial lesion wherein there is lactating adenoma and atypia. There are the presence of cystic ducts containing amorphous secretion [Figure 3] admixed with cellular debris, foamy histiocytes, and inflammatory infiltrates. The ducts are lined by ductal cells and with a tendency to show multi-layering. Most cells show secretory snouts. The ductal cells show large, ovoid, slightly pleomorphic nuclei with prominent nucleoli. Epithelial cells were not reactive to S100, and there was faint positive staining in the stroma and moderate stain on an active fibroblast [Figure 4].
Breast tissue shows a solid nodule mostly having hyalinized stroma [Figure 5] with a microscopic focus of necrotic cellular nest along the periphery. A larger focus of necrosis outside the nodule shows large, histiocyte-like cells having numerous, small, intracellular, round hyperchromatic bodies. The background shows abundant necrotic cellular debris and cells with reactive atypia. Fibroblasts in the stroma are immunoreactive to S100 while the epithelial cells are negative. There is also a faint reactivity in the stroma [Figure 6].
Breast tissue shows dilated ducts containing papillary structures [Figure 7] consisting of fibrovascular core superficially lined by two cell-type ductal epithelium. Some fibroblasts [Figure 8] are immunoreactive to S100 with. The epithelial cells were also immunonegative.
Breast tissue with overlying skin shows focal ulcer. The base of the ulcer shows a remnant gland or duct lined by multilayered atypical cells having large, ovoid, pleomorphic nuclei with prominent nucleoli and moderate, thick, eosinophilic cytoplasm [Figure 9]. These cells freely infiltrate the ulcer base and stroma and can be seen within endothelial-lined spaces. In immunostaining of S100, spindle-shaped cells (SP) [Figure 10] are immunoreactive with faint immunostaining in the stroma.
Spindle Cell Carcinoma
There is fibromuscular tissue with widespread infiltration of neoplastic spindly cells [Figure 11] occurring in random stream and whorls. These cells possess small to medium-sized ovoid, slightly pleomorphic nuclei and fibrillary, eosinophilic cytoplasm. In S100 immunostaining, SPs [Figure 12] are immunoreactive with faint staining in the stroma.
There is mesenchymal lesion and tumors made up of neoplastic chondroid [Figure 13] occurring in irregular islands with intervening highly cellular areas and pockets of necrosis. The cartilage cells and spindle-shaped cell are both immunoreactive to S100 [Figure 14].
Carcinoma with Neuroendocrine Differentiation
Breast tissue shows a mass made up of small, uniform, hyperchromatic cells in trabeculated nests separated by delicate fibroconnective tissue. Cells possess small, round, compact, hyperchromatic nuclei, and scant cytoplasm [Figure 15]. In S100 immunostaining, neuroendocrine cells and luminal epithelial cells are immunoreactive [Figure 16].
In H and E staining, breast tissue shows dilated ducts lined by multilayered neoplastic cells. The ducts contain abundant extracellular mucin within which float the neoplastic cells [Figure 17]. In S100 immunostaining, there is faint staining of some stromal cells [Figure 18]. No epithelial and myoepithelial cell immunoreactivity to S100 has been observed.
Invasive Ductal Carcinoma with Chondromyxoid Sarcoma or Carcinosarcoma
Breast tissue shows an admixture of ductal and chondromyxoid elements. The ductal cells which are carcinomatous possess large, ovoid, pleomorphic nuclei, and polygonal cytoplasmic occurring in solid nests with the glandular formation [Figure 19]. These carcinomatous cells are randomly distributed within abundant chondromyxoid stroma forming nodular masses [Figure 19]. In S100 immunostaining, some stromal cells were found to be positive [Figure 20].
There is proliferation of crowded tubular to complex glands separated by delicate and scanty fibroconnective intervening stroma [Figure 21]. Some glands form simple intertwined tuftings or short papillations while intraluminal bridging and fenestrations are seen in others. Myoepithelial cells are still evident. In S100 immunostaining, epithelial cells are negative while the stromal cells are highly immunoreactive [Figure 22].
For the controls used in the study, the negative control used is a section of skeletal muscle [Figure 23] and the positive control is a section from the skin [Figure 24]. These controls were used to indicate that the immunohistochemical procedure is optimized and working.
S100 has also been demonstrated in specific normal organs of other species such as in buffaloes,[7,9] sheep and goat, rats, poultry and rabbits, and humans. In these studies, the endothelial of capillaries, arteries, veins, and lymphatic vessels are regularly S100 protein immunoreactive. This is similarly observed in the endothelial cells of the blood vessels (BVs) in the normal lactating mammary and in the ten cases of CMTs wherein the endothelial cells of the BV particularly the arterioles are also immunoreactive with moderate to intense stain. Although normal BVs generally are not reactive with either monoclonal or polyclonal anti-S100 antibodies, weak labeling of a minority of endothelial cells are still noted. The immunoreactivity of these endothelial cells in both normal and tumorous mammary may be due to the regulatory activity of extracellular S100 proteins on the endothelial and vascular smooth muscle in order to participate in innate and adaptive immune responses, cell migration, and chemotaxis, tissue development and repair and leukocyte and tumor cell invasion. Since the endothelial cells of BVs in both normal and tumorous mammary are observed to positively react to S100, these cells cannot be used as markers for CMT.
There was also faint positive immunostaining observed on the milk secretions within the alveoli of a normal lactating mammary. This may imply the possible presence or expression of S100B in canine milk since S100B are normally found in small amounts in human milk.
The ten classifications of CMTs studied were all positive to S100 with notable differences in cells that were immunoreactive. The cells that are constantly observed moderately positive to S100 are the SPs and stromal cells as particularly observed in the majority of the obtained specimens, in cases of adenoma, fibroadenoma, ductal papillomatosis, complex carcinoma, spindle cell carcinoma, chondrosarcoma, mucinous carcinoma, carcinosarcoma, and tubular adenosis. These SPs and irregularly-shaped cells in the stroma are identified as fibrocytes and fibroblasts as confirmed in the visualization of these cells in the hematoxylin and eosin staining based on its morphology.
Fibrocytes are Commonly Reactive to S-100
Fibrocytes are the most common cells of the connective tissue or stroma. They are generally elongated and spindleshaped, with processes that contact adjacent cells and fibers, maintaining the connective tissue matrix by forming the fibers and constantly renewing the ground substance. On the other hand, fibroblasts have a larger, more euchromatic nucleus and more abundant, basophilic, cytoplasm than the fibrocyte and they are more active in the connective tissue matrix fibroblasts may arise directly from mesenchymal cells or are transformed from fibrocytes under the influence of microenvironmental factors such as cytokines. Fibroblasts have also been found to be a major cell type in the tumor stroma,[42,43] which was also observed in this study. Based on the study by Kalluri and Zeisberg, fibroblasts are associated with cancer cells at all stages of cancer progression and that their structural and functional contributions to this process are beginning to emerge. This suggests that production of growth factors, chemokines, and extracellular matrix by fibroblasts facilitates the angiogenic recruitment of certain cells. The impact of fibroblasts on tumor growth and progression has been the subject of the intensive investigation recently. As observed in this study, fibroblasts, fibrocytes and cartilage cells, which are cell types of mesenchymal origin, were positive to S100. The immunoreactivity of these cells to S100 may be explained by the expression of specific members of S100 in relation to their functions, particularly in mammary tumors. Some members of S100 protein family that have been noted in breast cancer are S100A4, S100A7, S100A8, S100A9, S100A6 and S100A11. These members of S100 have been said to exist as homodimers within cells and that upregulated gene expression of these proteins may imply the presence of a pathological condition such as tumor since overexpression of particular S100s has been found to be associated with tumorigenesis. One of the members of S100 is closely associated with fibroblasts, and that is the fibroblast-specific protein 1, also named as S100A4 which can be expressed by different cell types of mesenchymal origin. S100A4 expression found in the stroma had been found to contribute to metastatic dissemination. In human breast cancer cells, S100A4 overexpression is associated with increased migratory capacity since it was discovered that S100A4 has angiogenic effects. The explanation on this angiogenic effect of S100A4 is due to the interaction of this protein with Annexin II, and endothelial plasminogen co-receptor, and accelerated tPA-mediated plasminogen activation. This resulting local plasmin formation was found to contribute to tumor-induced angiogenesis and metastasis. Thus, both the angiogenic functions of fibroblasts and its expression of S100A4 visualized by immunoreactivity in S100 immunostaining may have an indication of an undergoing tumorigenesis in the tissue.
Chondrocytes are Moderately Reactive to S-100
Other cells that have been found to be moderately immunoreactive are chondrocytes, which were observed in the chondrosarcoma. These chondrocytes were positive to S100 probably due to its relation with fibroblasts as stated in a reference that in certain situations, fibroblasts may differentiate into chondroblasts which are active cartilage cells that form the matrix of cartilage and becomes chondrocytes when less inactive. Another possible explanation would be due to the other intracellular function of S100A4 at nanomolar levels, which stimulates matrix metalloproteinase 13 release from chondrocytes in a receptor for advanced glycation end products-mediated manner. Relatively, other members of S100, S100A8, and S100A9, also found in breast cancer, form a heterodimer complex implicated in regulating cell proliferation and in the metastatic process such as increasing the motility of cancer cells and facilitating the homing of migrating cells to pre-metastatic "niches" within the targets tissues.
Neuroendocrine Differentiation Reactivity to S-100
In one rare case, carcinoma with neuroendocrine differentiation, there were some neuroendocrine cells that were positive to S100. Neuroendocrine tumors of the breast are rare, accounting for <0.1% of all breast cancer and <1% of all neuroendocrine tumor. Neuroendocrine tumors are slow-growing tumors derived from neuroendocrine cells which are present throughout the body. These S 100-positive neuroendocrine cells may be due to the extracellular function of another member of S100 family, the S100B, which is expressed in astrocytes, Schwann cells, melanocytes, associate satellite cells, certain neuronal population and few other cell types. The positive immunoreaction of these neuroendocrine cells to S100 may be due to the sophisticated interrelationship of neuronal cells and glial cells.
Luminal Epithelium is Variably Reactive to S-100
Finally, luminal epithelial cells in the normal lactating canine mammary and in 9 CMTs were observed to be negative to S100 except for carcinoma with neuroendocrine differentiation wherein the epithelial cells were positive. This observation was parallel with the result in other a related study wherein some luminal epithelial cells were positive to S100 in some cases of human breast cancer.
This study was conducted to detect S100 immunohistochemically in CMT. In addition to the gold standard procedure, hematoxylin, and eosin staining, for histopathological diagnosis of tumors, the use of immunohistochemical staining was employed to determine the immunoreactivity of the different cells in CMT. 10 CMT samples were obtained from surgical resection in the different veterinary clinics and hospital in Metro Manila, Philippines, and a normal mammary sample was also obtained for comparison. The samples were processed for hematoxylin and eosin staining and S100 immunostaining. The hematoxylin and eosin stained sections were histopathologically diagnosed and classified according to the histological classifications of the mammary tumors of the dogs and cat. The malignant tumors were further graded using the Elston and Ellis method.
The S100 immunostaining of the sections showed that there is a moderate immunoreactive of the endothelial cells of the BVs in the normal lactating mammary and in the 10 cases of CMT. These cases of CMTs were all positive to S100. The cells that were found to be moderate to highly immunoreactive to S100 are SPs, chondrocytes, and stromal cells. While, in specific classification of CMT, carcinoma with neuroendocrine differentiation, some neuroendocrine cells were positive as well as in luminal epithelial cells. These positive staining of S100 in many cells was probably due to the intracellular and extracellular function of S100 in pathological conditions such as tumors.
S100 is detected in CMTs by the use of immunohistochemistry. The SPs, chondrocytes, and stromal cells are the cells that are immunoreactive to S100, therefore, these cells may possibly be used as markers for a tumor. In contrast, the endothelial cells of the BVs, which are positive both in normal mammary and in CMT, cannot be used as a marker. Although the ten CMTs are positive to S100, there are some differences regarding the cells that are positive. This difference is particular to a case, a carcinoma with neuroendocrine differentiation, wherein its luminal ductal epithelial cells and neuroendocrine cells are positive while those cells in other classifications of mammary tumors are negative. Thus, in the study of 10 classifications of CMT, immunohistochemical staining of S100 can be used as a marker in CMT.
[1.] Moore BW. A soluble protein characteristic of the nervous system. Biochem Biophys Res Commun 1965;19:739-44.
[2.] Schmitt FC, Bacchi CE. S-100 protein: Is it useful as a tumour marker in diagnostic immunocytochemistry? Histopathology 1989;15:281-8.
[3.] Donato R. S100: A multigenic family of calcium-modulated proteins of the EF-hand type with intracellular and extracellular functional roles. Int J Biochem Cell Biol 2001;33:637-68.
[4.] Kligman D, Hilt DC. The S100 protein family. Trends Biochem Sci 1988;13:437-43.
[5.] Maretta M, Marettova E. The localization of S100 protein in the bovine spleen. Folia Vet 2004;48:16-20.
[6.] Marettova E, Nad'ova P, Maretta M. Immunohistochemical localization of S-100 protein in the sheep spleen. Folia Vet 1998;42:19-22.
[7.] Vitor RJ, De Ocampo GD, Estacio MA. Immunohistochemical detection of S-100 in the spleen of the Philippine swamp buffalo (Bubalus bubalis carabanensis Castillo, 1998) (Artiodactyla: Bovidae). Philipp J Vet Med 2013;50:1-6.
[8.] Maretta M, Marettova E. Immunohistochemical demonstration of vimentin and S-100 protein in the kidneys. Gen Physiol Biophys 1999;18 Suppl 1:100-2.
[9.] Cena RB, Vitor RJ, De Ocampo GD, Estacio MA. Immunohistochemical detection of S100 in the kidney of the Philippine swamp buffalo (Bubalus bubalis carabanensis Castillo, 1998) (Artiodactyla: Bovidae). Philipp J Vet Med 2015;52:65-70.
[10.] Molin SO, Rosengren L, Baudier J, Hamberger A, Haglid K. S-100 alpha-like immunoreactivity in tubules of rat kidney. A clue to the function of a "brain-specific" protein. J Histochem Cytochem 1985;33:367-74.
[11.] de Girolamo P, Arcamone N, Gargiulo G. S 100-like protein in the goldfish (Carassius auratus) kidney. An immunohistochemical study. Cell Tissue Res 2000;302:135-8.
[12.] De Girolamo P, Arcamone N, Pelagalli GV, Gargiulo G. Immunohistochemical localization of S 100-like protein in non-mammalian kidney. Microsc Res Tech 2003;60:652-7.
[13.] Abd-Elmaksoud A, Shoeib MB, Marei HE. Localization of S-100 proteins in the testis and epididymis of poultry and rabbits. Anat Cell Biol 2014;47:180-7.
[14.] Cruzana BC, Hondo E, Kitamura N, Matsuzaki S, Nakagawa M, Yamada J, et al. Differential localization of immunoreactive alpha- and beta-subunits of S-100 protein in feline testis. Anat Histol Embryol 2000;29:83-6.
[15.] Hanaue M, Miwa N, Takamatsu K. Immunohistochemical characterization of S100A6 in the murine ovary. Acta Histochem Cytochem 2012;45:9-14.
[16.] McLaren KM, Cossar DW. The immunohistochemical localization of S100 in the diagnosis of papillary carcinoma of the thyroid. Hum Pathol 1996;27:633-6.
[17.] Takashi M, Haimoto H, Murase T, Mitsuya H, Kato K. An immunochemical and immunohistochemical study of S100 protein in renal cell carcinoma. Cancer 1988;61:889-95.
[18.] Sandusky GE Jr., Carlton WW, Wightman KA. Immunohistochemical staining for S100 protein in the diagnosis of canine amelanotic melanoma. Vet Pathol 1985;22:577-81.
[19.] Donato R, Cannon BR, Sorci G, Riuzzi F, Hsu K, Weber DJ, et al. Functions of S100 proteins. Curr Mol Med 2013;13:24-57.
[20.] Destexhe E, Lespagnard L, Degeyter M, Heymann R, Coignoul F. Immunohistochemical identification of myoepithelial, epithelial, and connective tissue cells in canine mammary tumors. Vet Pathol 1993;30:146-54.
[21.] Sleeckx N, de Rooster H, Veldhuis Kroeze EJ, Van Ginneken C, Van Brantegem L. Canine mammary tumours, an overview. Reprod Domest Anim 2011;46:1112-31.
[22.] Moe L. Population-based incidence of mammary tumours in some dog breeds. J Reprod Fertil Suppl 2001;57:439-43.
[23.] Egenvall A, Bonnett BN, Ohagen P, Olson P, Hedhammar A, von Euler H. Incidence of and survival after mammary tumors in a population of over 80,000 insured female dogs in Sweden from 1995 to 2002. Prev Vet Med 2005;69:109-27.
[24.] DeSantis C, Ma J, Bryan L, Jemal A. Breast cancer statistics, 2013. CA Cancer J Clin 2014;64:52-62.
[25.] Misdorp W, Cotchin E, Hampe JF, Jabara AG, von Sandersleben J. Canine malignant mammary tumours I. Sarcomas. Vet Pathol 1971;8:99-117.
[26.] Misdorp W, Cotchin E, Hampe JF, Jabara AG, Sandersleben JV Canine malignant mammary tumours II. adenocarcinomas, solid carcinomas and spindle cell carcinomas. Vet Pathol 1972;9:447-70.
[27.] Misdorp W, Cotchin E, Hampe JF, Jabara AG, Sandersleben JV. Canine Malignant Mammary Tumors III. Special Types of Carcinomas, Malignant Mixed Tumors. Vet Pathol. 1973;10(3):241-56.
[28.] Misdorp W, Else, RW, Hellmen, E, Lipscomb, TP. Histological Classification of Mammary Tumors of the Dog and the Cat. Washington, DC: Armed Forces Institute of Pathology; 1999.
[29.] Shafiee G, Mohajeri-Tehrani M, Pajouhi M, Larijani B. The importance of hypoglycemia in diabetic patients. J Diabetes Metab Disord 2012;11:17.
[30.] Hsu SM, Raine L, Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: A comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 1981;29:577-80.
[31.] Flens MJ, van der Valk P, Tadema TM, Huysmans AC, Risse EK, van Tol GA, et al. The contribution of immunocytochemistry in diagnostic cytology. Comparison and evaluation with immunohistology. Cancer 1990;65:2704-11.
[32.] Hsieh HL, Schafer BW, Sasaki N, Heizmann CW. Expression analysis of S100 proteins and RAGE in human tumors using tissue microarrays. Biochem Biophys Res Commun 2003;307:375-81.
[33.] von Schoultz E, Hansson LO, Djureen E, Hansson J, Karnell R, Nilsson B, et al. Prognostic value of serum analyses ofS-100 beta protein in malignant melanoma. Melanoma Res 1996;6:133-7.
[34.] Sedaghat F, Notopoulos A. S100 protein family and its application in clinical practice. Hippokratia 2008;12:198-204.
[35.] Ohsie SJ, Sarantopoulos GP, Cochran AJ, Binder SW. Immunohistochemical characteristics of melanoma. J Cutan Pathol 2008;35:433-44.
[36.] Bhatt AN, Mathur R, Farooque A, Verma A, Dwarakanath BS. Cancer biomarkers-current perspectives. Indian J Med Res 2010;132:129-49.
[37.] Elston CW, Ellis IO, editors. The Breast. 3rd ed. London: Churchill Livingstone; 1998.
[38.] Parkkila S, Pan PW, Ward A, Gibadulinova A, Oveckova I, Pastorekova S, et al. The calcium-binding protein S100P in normal and malignant human tissues. BMC Clin Pathol 2008;8:2.
[39.] Wick M. Monoclonal Antibodies in Diagnostic Immunohistochemistry. England: Taylor & Francis; 1988.
[40.] Gazzolo D, Monego G, Corvino V, Bruschettini M, Bruschettini P, Zelano G, et al. Human milk contains S100B protein. Biochim Biophys Acta 2003;1619:209-12.
[41.] Dellman HD, Eurell JA. Textbook of Veterinary Histology. 5th ed. Philadelphia: Lippincott Williams & Wilkins; 1998.
[42.] Kalluri R, Zeisberg M. Fibroblasts in cancer. Nat Rev Cancer 2006;6:392-401.
[43.] Li J, Chen L, Qin Z. Multifaceted tumor stromal fibroblasts. Cancer Microenviron 2012;5:187-93.
[44.] Zhang J, Chen L, Liu X, Kammertoens T, Blankenstein T, Qin Z, et al. Fibroblast-specific protein 1/S100A4-positive cells prevent carcinoma through collagen production and encapsulation of carcinogens. Cancer Res 2013;73:2770-81.
[45.] Strutz F, Okada H, Lo CW, Danoff T, Carone RL, Tomaszewski JE, et al. Identification and characterization of a fibroblast marker: FSP1. J Cell Biol Nat Cell Biol 2006;8:1321-3.
[46.] Bresnick AR, Weber DJ, Zimmer DB. S100 proteins in cancer. Nat Rev Cancer 2015;15:96-109.
[47.] Ambartsumian N, Klingelhofer J, Grigorian M, Christensen C, Kriajevska M, Tulchinsky E, et al. The metastasis-associated mts1(S100A4) protein could act as an angiogenic factor. Oncogene 2001;20:4685-95.
[48.] Semov A, Moreno MJ, Onichtchenko A, Abulrob A, Ball M, Ekiel I, et al. Metastasis-associated protein S100A4 induces angiogenesis through interaction with annexin II and accelerated plasmin formation. J Biol Chem 2005;280:20833-41.
[49.] Donato R. RAGE: A single receptor for several ligands and different cellular responses: The case of certain S100 proteins. Curr Mol Med 2007;7:711-24.
[50.] Rafii S, Lyden D. S100 chemokines mediate bookmarking of premetastatic niches. Nat Cell Biol 2006;8:1321-3.
[51.] Ogawa H, Nishio A, Satake H, Naganawa S, Imai T, Sawaki M, et al. Neuroendocrine tumor in the breast. Radiat Med 2008;26:28-32.
[52.] Singh S, Aggarwal G, Kataria SP, Kalra R, Duhan A, Sen R. Primary neuroendocrine carcinoma of breast. J Cytol 2011;28:91-2.
[53.] Morale MC, Gallo F, Tirolo C, Testa N, Caniglia S, Marletta N, et al. Neuroendocrine-immune (NEI) circuitry from neuron-glial interactions to function: Focus on gender and HPA-HPG interactions on early programming of the NEI system. Immunol Cell Biol 2001;79:400-17.
[54.] Hein SM, Haricharan S, Johnston AN, Toneff MJ, Reddy JP, Dong J, et al. Luminal epithelial cells within the mammary gland can produce basal cells upon oncogenic stress. Oncogene 2016;35:1461-7.
Khristine Kaith S Lloren (1), Rodel Jonathan S Vitor II (2)
(1) College of Veterinary Medicine and Agricultural Sciences, De La Salle Araneta University, Malabon, Metro Manila, Philippines, (2) Department of Biology, College of Science, De La Salle University, Manila, Philippines
Correspondence to: Rodel Jonathan Santos Vitor II, E-mail: email@example.com
Received: February 07, 2018; Accepted: February 24, 2018
How to cite this article: Lloren KKS, Vitor RJS. S100 can be used as a tumor marker in canine mammary tumors. Natl J Physiol Pharm Pharmacol 2018;8:928-939.
Source of Support: Nil, Conflict of Interest: None declared.
Table 1: Classification (according to WHO) of 10 CMT Type of tumor n Benign tumors 3 Simple adenoma 1 Fibroadenoma 1 Ductal papillomatosis 1 Malignant tumors 6 Complex carcinoma 1 Spindle cell carcinoma 1 Chondrosarcoma 1 Carcinoma 1 Mucinous carcinoma 1 Carcinosarcoma 1 Hyperplasia and dysplasia 1 Adenosis 1 WHO: World Health Organization, CMT: Canine mammary tumors Table 2: Immunohistochemical reactivity of cell types for S100 in 10 CMTs and in normal lactating mammary gland Type of tumor Cell types or structures and immunoreactivity Positive Negative Normal lactating mammary gland En E, R, P SP Benign tumors Simple adenoma En, SC E, R, P Fibroadenoma En, SP, SC E, R, P Ductal papillomatosis En, SP E, R, P Malignant tumors Complex carcinoma En, SC E, R, P Spindle cell carcinoma En, SP E, R, P Chondrosarcoma En, SP, C E, R, P Carcinoma with endocrine En, E, NEC S, R, P differentiation Mucinous carcinoma En, SC E, R, P Carcinosarcoma En, SC E, R, P Hyperplasia and dysplasia Adenosis En, SC E, R, P SP: Spindle-shaped cells, C: Cartilage cells, SC: Stromal cells, E: Epithelial alveolar or ductal cells, R: Resting myoepithelial cells, P: Proliferative suprabasal myoepithelial cells, En: Endothelial cell of blood vessel, NEC: Neuroendocrine cells. CMTs: Canine mammary tumors
|Printer friendly Cite/link Email Feedback|
|Title Annotation:||RESEARCH ARTICLE|
|Author:||Lloren, Khristine Kaith S.; Vitor, Rodel Jonathan S., II|
|Publication:||National Journal of Physiology, Pharmacy and Pharmacology|
|Date:||Dec 15, 2018|
|Previous Article:||Assignment-based learning of essential medicine list concepts among undergraduate medical students.|
|Next Article:||A study on antidepressant-like effect of dihydroxy flavones in mice and their mechanisms involved.|