High incidence of chromosome 1 abnormalities in a series of 27 renal oncocytomas: cytogenetic and fluorescence in situ hybridization studies.

* Context.--It has recently been shown by cytogenetics that there is a high incidence of chromosome 1 abnormalities in renal oncocytomas.

Objective.--To confirm the cytogenetic results by fluorescence in situ hybridization (FISH) analysis.

Design.--Nine additional cytogenetic analyses were added to those reported in our recent study, with a total of 27 tumors studied, which makes it the largest series of renal oncocytomas studied to date by cytogenetics and/or FISH. We used the LSI 1p36/LSI 1q25 Dual Color Probe Set to make the analyses.

Results.--In this study, combined cytogenetics and FISH showed loss of chromosome arm 1p1 in 48% of renal oncocytomas. By FISH, deletion of 1p36.3 was observed in 59% of renal oncocytomas, whereas by cytogenetics, abnormality in chromosome 1 was seen in 32% of tumors. However, the incidence of chromosome 1 abnormalities among 9 bilateral tumors was much higher than in single tumors (88% vs 28%, respectively). Loss of only the 1p36.3 site occurred in 2 renal oncocytomas with translocation of chromosome 1, as shown by cytogenetics. Concordance between the 2 techniques, when they were used simultaneously to detect chromosome 1p1 abnormality, was 82%.

Conclusions.--This study further confirmed our prior results demonstrating the widespread occurrence of chromosome 1 abnormalities in renal oncocytomas. Although no abnormalities in chromosome 1 in tumors with normal karyotypes were detected by FISH using the current set of probes, a much higher incidence of such abnormalities was found in bilateral tumors, suggesting that genetic alterations related to the development of renal oncocytoma reside in this region.

(Arch Pathol Lab Med. 2007;131:81-85)

**********

Approximately 3% to 5% of renal tumors are classified histologically as oncocytoma, a benign tumor characterized by abundant granular eosinophilic cytoplasm rich in mitochondria and believed to arise from the intercalated cells of the collecting ducts. (1-4) Histologically, renal oncocytoma may mimic eosinophilic variants of chromophobe renal cell carcinoma (ChRCC), clear cell carcinoma, and type 2 papillary renal cell carcinoma.

According to the Mitelman Database of Chromosome Aberrations in Cancer (2006), (5) 75 oncocytomas have been analyzed to date. Cytogenetic studies showed heterogeneity among oncocytomas: some tumors were shown to have a normal karyotype, but a subset of oncocytomas showed loss of chromosome 1 or the short arm of chromosome 1, loss of the Y chromosome (in male patients), and occasionally, monosomy of chromosome 14; a few tumors were reported to have structural rearrangements involving chromosome 11 band q13. (6-13) Abnormality in chromosome 1 was also present in ChRCC, which has given rise to the hypothesis that the subset of oncocytomas with this abnormality may be related to ChRCC. In contrast, no chromosome 1 abnormalities are found in clear cell or type 2 papillary renal cell carcinoma. (3)

Using conventional cytogenetics in a series of 13 histologically well-defined oncocytomas, we recently reported that there is a high incidence of chromosome 1 abnormality in oncocytoma. (12) Because cytogenetic analysis is time-consuming and tedious, and fresh tissue is not always available, we sought to confirm the cytogenetic results by FISH analysis in this study. Additional cases have been included, with a total of 27 tumors studied, which makes it the largest series of oncocytomas studied to date by cytogenetic and/or FISH methods. We also sought to test whether renal oncocytomas with cytogenetically normal chromosome 1 have abnormalities in the 1p36.3 locus.

MATERIALS AND METHODS

Tissue Specimens

A total of 27 renal oncocytomas from 22 patients who underwent nephrectomy at Loyola University Medical Center from 1996 to 2003 were included in the study. This study was approved by the institutional review board. There were 12 male and 10 female patients with ages ranging from 42 to 85 years (mean, 65; median, 69). The histopathologic diagnosis of renal oncocytoma was based on the 2003 World Health Organization classification of renal epithelial tumors (3) and specifically included histochemistry (Hale colloidal iron stain), immunohistochemistry (cytokeratin 7, CD10, vimentin), and electron microscopy as previously described. (14) Two tumors (cases 12 and 13 in the Table) showed hybrid morphology with focal areas (approximately 25%) of cells with cytoplasmic membrane condensation and prominent perinuclear clearing (resembling eosinophilic variant of chromophobe-like cells) in a background of classic oncocytic cells. In these 2 tumors there was focal reticular cytoplasmic staining with Hale colloidal iron and focal positivity for cytokeratin 7. All other tumors were negative for cytokeratin 7 and Hale colloidal iron, and all tumors were negative for vimentin and CD10. (1-4,12) Electron microscopy demonstrated abundant mitochondria and no microvesicles in all 21 tumors studied. (4) The renal oncocytomas were bilateral in 4 cases and unilateral in 18 cases, plus 1 additional tumor was sampled from multifocal disease (total of 27 tumors). One patient (case 3) had multiple microscopic renal oncocytoma foci in addition to bilateral renal involvement (renal oncocytosis). None of the 22 patients had a family history of concurrent renal oncocytomas. Analysis by FISH was performed in 22 of 27 renal oncocytomas and the results of conventional cytogenetics were available in 22 of 27 tumors. The results of cytogenetic analysis of 13 of these 22 renal oncocytomas have been previously reported. (12) The results of combined FISH and conventional cytogenetics were available in 17 of 27 renal oncocytomas, allowing correlation of results.

In Situ Hybridization 

Sections at 5 [micro]m thickness were prepared from paraffin-embedded tissues. A LSI 1p36/LSI 1q25 Dual Color Probe Set from Vysis/Abbott Molecular, Inc (Des Plains, Ill) was used for analysis. This comprised a mixture of ~400 kb SpectrumOrange (Abbott) -labeled 1p36 sequence and ~620 kb SpectrumGreen (Abbott) -labeled 1q25 control probe. The 1p36 probe contained sequences extending from near the SHGC57243 locus through the TP73 and EGFL3 genes, and ending proximally telomeric to the EGFL3 and RH75821 loci. The 1q25 probe contains sequences extending from a point telomeric to the WI-6848 locus, through the ABL2 and ANGPTL1 genes, and ending proximally near SHGC-1322. In situ hybridization was performed using the Vysis/Abbott paraffin pretreatment II procedure except that protease solution of 2 mg/mL was used.

Evaluation of FISH Signals

The fluorescence signals were scored by three observers (K.J., G.P.P., Y.C.). At least 30 nonoverlapping interphase cell nuclei were scored per observer in each tumor. In 13 cases, nuclei of nonneoplastic kidney tubular cells from the adjacent normal parenchyma were also evaluated as a negative control, and 10 cases of ChRCC were included as positive controls. The cutoff for deletion was set as a mean per cell count of 1.5 or less. A mean per cell count of 2.5 or greater was considered trisomic.

Conventional Cytogenetics

Cytogenetic analyses were performed as previously described. (12)

RESULTS

The combined FISH and conventional cytogenetic results in renal oncocytomas are summarized in the Table. Overall, loss of chromosome arm 1p1 was seen in 13 (48%) of 27 tumors. No abnormality of chromosome 1 was detected by either method in 13 (48%) of 27 tumors; in 1 case (case 18) studied by FISH alone, trisomy of chromosome 1 was seen.

Among 22 tumors studied by FISH analysis, deletion of 1p36.3 was observed in 13 (59%) of the tumors. In 11 (85%) of 13 tumors, deletion of 1p36.3 was accompanied by deletion of 1q25, indicating monosomy 1 (Figure, A), whereas in 2 (15%) of 13 tumors there was an isolated deletion of 1p36.3 (Figure, B). Both signals for 1p36.3 and 1q25 were seen in 8 (36%) of 22 tumors, indicating disomy (Figure, C). Among 8 bilateral tumors studied by FISH, all showed deletion of 1 signal each of 1p36.3 and 1q25 (see following discussion also). Three signals for 1p36.3/ 1q25 were observed in 1 tumor. All of the adjacent non-neoplastic renal tubular cells were disomic for 1p36.3/ 1q25 and cases of ChRCC showed loss of chromosome 1 (data not shown).

[FIGURES A-C OMITTED]

Among 22 tumors studied by conventional cytogenetics, abnormality of chromosome 1 was seen in 7 (32%) tumors. Five (71%) of 7 tumors showed loss of the entire chromosome 1. In 2 (29%) of 7 tumors, there was loss of 1p as a result of unbalanced translocation [der(1;13)(q10;q10) and der(1;5)(q10;p10)].

Both FISH and conventional cytogenetic results were available for comparison in 17 tumors. There was agreement of results in 14 (82%) of 17 cases. Loss of chromosome arm 1p1 was concurrently observed with both methods in 7 (41%) of 17 tumors. All 5 tumors with chromosome 1 monosomy by cytogenetics (cases 1, 2 [right and left], 5, and 6) showed concordant 1p36.31q25 loss by FISH. The 2 tumors with chromosome 1p loss by translocation (cases 9 and 10) also showed loss of 1p36.3 only by FISH analysis. The 3 discordant tumors (cases 3 [right], 4 [left, site 1], and 7) showed loss of 1p36.31p25 by FISH with no abnormality of chromosome 1 by conventional cytogenetic analysis. In contrast, however, all 7 tumors with disomy in 1p36.31q25 by FISH (cases 7 and 11-16) correspondingly showed normal chromosome 1 by cytogenetic analysis. One tumor showed apparent trisomy of chromosome 1 by FISH; however, no cytogenetic data were available for comparison.

There were 9 bilateral tumors, of which 8 were studied by FISH and 6 by cytogenetics (cases 1-4). Overall in this group, 8 (89%) of 9 tumors showed loss of chromosome 1 by at least one of the methods. By comparison, among the 18 unilateral tumors studied by cytogenetics and/or FISH, loss of chromosome arm 1p1 was seen in 5 (28%) of the tumors.

COMMENT

Prior cytogenetic studies have detected a varying incidence of chromosome arm 1p1 loss in renal oncocytomas ranging from 0% to 75%. (4-13) In contrast, prior FISH studies reported this frequency as ranging from 10% to 23%. (15,16) This varying incidence of chromosome 1 involvement may be the result of interobserver variation in the histopathologic diagnosis of renal oncocytoma and/or the use of different genetic techniques. Our study, which is the largest to date, was based on a series of histopathologically well-defined oncocytomas with the diagnoses supported by immunohistochemistry, colloidal iron stain, and electron microscopic studies. (1-4,12) Our studies show a higher incidence of chromosome 1 abnormalities in oncocytomas than has been reported thus far in the literature. (5-13,15,16) By combined analysis of interphase and metaphase cells, we demonstrated significant loss of chromosome arm 1p1 in renal oncocytomas (48%). We interpret this discordance between our data and the prior reports of others as a consequence of differences in methods. Brown et al (16) and Brunelli et al (15) used different chromosome 1 probes (proximal 1q [1qh] region and centromeric probes, respectively), which would detect monosomy only. In contrast, the set of probes that we used has the ability to detect both monosomy 1 and loss of a small portion of 1p36.3.

Interestingly, our combined cytogenetic and FISH studies are concordant with results emerging from recent comparative genomic hybridization studies, in which 49% of renal oncocytomas were reported to show losses at 1p. (13,17) In our study, the incidence of chromosome 1 abnormalities in bilateral tumors was much higher than in single tumors (88% vs 28%, respectively). Prior studies also reported a higher incidence of cytogenetic abnormalities in bilateral/ synchronous oncocytomas. (18-20) A much higher incidence of chromosome 1 abnormalities among bilateral tumors (88%) suggests that this chromosome is important in genesis of renal oncocytomas.

We observed a good correlation between FISH and conventional cytogenetics of 82%. All of the renal oncocytomas included in our study that showed loss of chromosome arm 1p1 by conventional cytogenetics also showed a corresponding loss by FISH analysis, which validates the selection of this particular set of probes. In 2 tumors in our series, cytogenetics demonstrated loss of 1p only (by translocation), and we also observed loss of 1p36.3 only by FISH. Interestingly, in 3 tumors, FISH detected losses in tumors that had normal chromosome 1 by conventional cytogenetics. This observation of a higher incidence of chromosomal aberrations detected by FISH than by conventional cytogenetics is not unique to our study. (21) The possibility that the cytogenetic cell cultures may have been overgrown by stromal cells is a probable explanation. Nonrandom clonal chromosome arm 1p1 loss has not been reported to occur in nonneoplastic kidney tissue. (22,23)

In a single tumor we detected trisomy of chromosome 1 by FISH; unfortunately, no cytogenetic data are available for comparison. Although chromosomal gains have been seen on occasion in oncocytoma (including the current series), we are not aware of reports of chromosome 1 gains in oncocytoma, and further studies are needed to address this issue.

Van der Berg et al (10) proposed that a subset of renal oncocytomas with loss of chromosome 1 may be related to ChRCCs because the latter also show loss of chromosome 1 (as well as multiple additional monosomies of 2, 6, 10, 13, 17, 21, and X, or loss of Y in male patients). Morphologic (including ultrastructural studies), cytogenetic, and FISH studies appear to support the concept of a morphologic and biologic continuum in at least a subset of these tumors, with some tumors showing clearly "hybrid" features. (4,10,12,20,24) In our series, some of the oncocytomas showed a complex karyotype (chromosome 21 loss and an isochromosome 9p with 9q loss). It has been proposed that multiple chromosomal losses, including that of chromosome 21, may result in progression of oncocytoma to chromophobe carcinoma. (4,10,12,20,24) Recently, Fuzesi et al (13) proposed that del(14q) may represent distinct cytogenetic aberration in a subset of renal oncocytomas. We previously reported abnormalities in chromosome 14 and these are again seen in several tumors in the current series. (12) Thus, heterogeneity among oncocytomas is being further supported by these more recent studies.

The issue of the tumors with nonrecurrent or no detectable abnormalities by cytogenetic studies remains an open question. Thus far, using the present methods, we have not observed any isolated deletion of 1p36.3 in tumors in which both chromosomes 1 were normal by conventional cytogenetics. Although prior studies have suggested that a tumor suppressor gene important for the genesis of renal oncocytomas might reside on chromosome 1p, this gene has not yet been identified. (25,26) The subtelomeric portion of chromosome 1 has the highest concentration of genes, and the 1p36 locus contains the gene encoding p73, a protein with considerable homology to the p53 tumor suppressor. (27) Deletion of the 1p36 region has frequently been observed in nonrenal tumors, including oligodendroglioma, ovarian carcinoma, malignant melanoma, and neuroblastoma. (28-31) Thus, additional molecular studies of this interesting region may shed light on the location of the putative tumor suppressor gene involved in the genesis of renal oncocytomas.

In conclusion, using a combination of interphase FISH and metaphase cytogenetic analysis, we demonstrated widespread occurrence of loss of chromosome arm 1p1 and a high concordance between these 2 techniques in detecting this abnormality. In this study, FISH had a greater level of detection of chromosome 1 abnormalities than cytogenetics. Although distinction between oncocytoma and ChRCC represents a more complex issue, demonstration of loss of chromosome arm 1p1 may be helpful in differentiating renal oncocytomas from the eosinophilic variants of clear cell and papillary renal cell carcinoma, especially in small biopsy samples. Location of the tumor suppressor gene candidate requires further studies, but a much higher incidence of chromosome 1 abnormalities in bilateral tumors suggests that this chromosome is important in genesis of renal oncocytomas.

Accepted for publication June 9, 2006.

References

(1.) Amin MB, Crotty TB, Tickoo SK, Farrow GM. Renal oncocytoma: a reappraisal of morphologic features with clinicopathologic findings in 80 cases. Am J Surg Pathol. 1997;21:1-12.

(2.) Perez-Ordonez B, Hamed G, Campbell S, et al. Renal oncocytoma: a clinicopathologic study of 70 cases. Am J Surg Pathol. 1997;21:871-883.

(3.) Reuter VE, Davis CJ, Moch H. Oncocytoma. In: Pathology and Genetics of Tumours of the Urinary System and Male Genital Organs. Lyon, France: IARC Press; 2004:9-87. World Health Organization Classification of Tumours; vol 6.

(4.) Picken MM. The evolving concept of renal neoplasia: impact of emerging molecular and electron microscopic studies. Ultrastruct Pathol. 2005;29:277-282.

(5.) Mitelman F, Johansson B, Mertens F, eds. Mitelman Database of Chromosome Aberrations in Cancer (2006). Available at: http://cgap.nci.nih.gov/Chromosomes/ Mitelman. Accessed January 8, 2006.

(6.) Fuzesi L, Gunawan B, Braun S, et al. Cytogenetic analysis of 11 renal oncytomas: further evidence of structural rearrangements of 11q13 as a characteristic chromosomal anomaly. Cancer Genet Cytogenet. 1998;107:1-6.

(7.) Kovacs G, Szucs S, Eichner W, Maschek HJ, Wahnschaffe U, De Riese W. Renal oncocytoma: a cytogenetic and morphologic study. Cancer. 1987;59:2071- 2077.

(8.) Dobin SM, Harris CP, Reynolds JA, et al. Cytogenetic abnormalities in renal oncocytic neoplasms. Genes Chromosomes Cancer. 1992;4:25-31.

(9.) Crotty TB, Lawrence KM, Moertel CA, et al. Cytogenetic analysis of six renal oncocytomas and a chromophobe renal cell carcinoma: evidence that -Y, -1 may be a characteristic anomaly in renal oncocytomas. Cancer Genet Cytogenet. 1992;61:61-66.

(10.) Van den Berg E, Dijkhuizen T, Storkel S, et al. Chromosomal changes in renal oncocytomas: evidence that t(5;11)(q35;q13) may characterize a second subgroup of oncocytomas. Cancer Genet Cytogenet. 1993;79:164-168.

(11.) Jhang JS, Narayan G, MurtyVV, Mansukhani MM. Renal oncocytomas with 11q13 rearrangements: cytogenetic, molecular and immunohistochemical analysis of cyclin D1. Cancer Genet Cytogenet. 2004;149:114-119.

(12.) Lindgren V, Paner GP, Omeroglu A, et al. Cytogenetic analysis of a series of 13 renal oncocytomas. J Urol. 2004;171:602-604.

(13.) Fuzesi L, Frank D, Nguyen C, Ringert R, Bartels H, Gunawan B. Losses of 1p and chromosome 14 in renal oncocytomas. Cancer Genetic Cytogenet. 2005; 160:120-125.

(14.) Picken MM, Curry JL, Lindgren V, Clark JI, Eble JN. Metanephric adenosarcoma in a young adult: morphologic, immunophenotypic, ultrastructural, and fluorescence in situ hybridization analyses: a case report and review of the literature. Am J Surg Pathol. 2001;25:1451-1457.

(15.) Brunelli M, Eble JN, Zhang S, Martignoni G, Delahunt B, Cheng L. Eosinophilic and classic chromophobe renal cell carcinomas have similar frequent losses of multiple chromosomes from among chromosomes 1, 2, 6, 10, and 17, and this pattern of genetic abnormality is not present in renal oncocytoma. Mod Pathol. 2005;18:161-169.

(16.) Brown JA, Takahashi S, Alcaraz A, et al. Fluorescence in situ hybridization analysis of renal oncocytoma reveals frequent loss of chromosomes Y and 1. J Urol. 1996;156:31-35.

(17.) Presti JC, Moch H, Reuter VE, Huynh D, Waldman FM. Comparative genomic hybridization for genetic analysis of renal oncocytomas. Genes Chromosomes Cancer. 1996;17:199-204.

(18.) Dal Cin P, Van Poppel H, Van Damme B, Baert L, Van den Berghe H. Cytogenetic investigation of synchronous bilateral renal tumors. Cancer Genet Cytogenet. 1996;89:57-60.

(19.) Al-Saleem T, Cairns P, Dulaimi EA, Feder M, Testa JR, Uzzo RG. The genetics of renal oncocytosis: a possible model for neoplastic progression. Cancer Genet Cytogenet. 2004;152:23-28.

(20.) Pavlovich CP, Grubb RL III, Hurley K, et al. Evaluation and management of renal tumors in the Birt-Hogg-Dube syndrome. J Urol. 2005;173:1482-1486.

(21.) Wada Y, Yokogi H, Moriyama-Gonda N, Shigeno K, Shiina H, Igawa M. Chromosome aberrations in renal tumors detected by fluorescence in situ hybridization. Cancer Genet Cytogenet. 1997;99:38-44.

(22.) Casalone R, Granata Casalone P, Minelli E, et al. Significance of the clonal and sporadic chromosome abnormalities in non-neoplastic renal tissue. Hum Genet. 1992;90:71-78.

(23.) Van den Berg E, Dijkhuizen T, Storkel S, Molenaar WM, de Jong B. Chromosomal abnormalities in non-neoplastic renal tissue. Cancer Genet Cytogenet. 1995;85:152-154.

(24.) Psihramis KE, Dal Cin P, Dretler SP, Prout GR Jr, Sandberg AA. Further evidence that renal oncocytoma has malignant potential. J Urol. 1988;139:585- 587.

(25.) Trash-Bingham CA, Salazar H, Greenberg RE, Tartof KD. Loss of heterozygosity studies indicate that chromosome arm 1p harbors a tumor suppressor gene for renal oncocytomas. Genes Chromosomes Cancer. 1996;16:64-67.

(26.) Feder M, Liu Z, Apostolou S, Greenberg RE, Testa JR. Loss of chromosomes 1 and X in a renal oncocytoma: implications for a possible pseudoautosomal suppressor locus. Cancer Genet Cytogenet. 2000;123:71-72.

(27.) Saccone S, De Sario A, Della Valle G, Bernardi G. The highest gene concentrations in the human genome are in telomeric bands of metaphase chromosomes. Proc Natl Acad Sci U S A. 1992;89:4913-4917.

(28.) Ragnarsson G, Eiriksdottir G, Johannsdottir JT, Jonasson JG, Egilsson V, Ingvarsson S. Loss of heterozygosity at chromosome 1p in different solid human tumours: association with survival. Br J Cancer. 1999;79:1468-1474.

(29.) Thompson FH, Taetle R, Trent JM, et al. Band 1p36 abnormalties and t(1; 17) in ovarian carcinoma. Cancer Genet Cytogenet. 1997;96:106-110.

(30.) Nelson MA, Ariza ME, Yang JM, et al. Abnormalities in the p34cdc2-related PITSLRE protein kinase gene complex (CDC2L) on chromosome band 1p36 in melanoma. Cancer Genet Cytogenet. 1999;108:91-99.

(31.) Kaghad M, Bonnet H, Yang A, et al. Monoallelically expressed gene related to p533 at 1p36, a region frequently deleted in neuroblastoma and other human cancers. Cell. 1997;90:809-819.

Gladell P. Paner, MD; Valerie Lindgren, PhD; Kris Jacobson, MS; Kathleen Harrison, PhD; Ying Cao, MD, PhD; Steve C. Campbell, MD, PhD; Robert C. Flanigan, MD; Maria M. Picken, MD, PhD

From the Departments of Pathology (Drs Paner, Lindgren, Harrison, Cao, and Picken) and Urology (Drs Campbell and Flanigan), Loyola University Medical Center, Maywood, Ill; and Research and Development, Vysis/Abbott Molecular, Inc, Des Plains, Ill (Ms Jacobson). Dr Lindgren is currently with the University of Illinois in Chicago, Department of Pathology. Dr Campbell is currently with the Cleveland Clinic, Glickman Urological Institute, Cleveland, Ohio.

Previously presented in part at the 93rd Annual Meeting of the United States and Canadian Academy of Pathology in Vancouver, British Columbia, March 2004.

Ms Jacobson is an employee of Vysis/Abbott Molecular, Inc. All other authors have no relevant financial interest in the products or companies described in this article.

Reprints: Maria M. Picken, MD, PhD, Department of Pathology, LUMC, Bldg 110, Room 2242, 2160 S First Ave, Maywood, IL 60153 (e-mail: [e-mail]).

Interphase Fluorescence In Situ Hybridization (FISH) Analysis of
Chromosome 1 and Corresponding Karyotype by Conventional G-Banding
Technique of Renal Oncocytomas *

                                                 FISH Analysis

Case                Side         Size, cm       1p36.3       1q25

 1 ([dagger])   Right              12.2        Monosomy    Monosomy
                Left                5.0        Monosomy    Monosomy
 2 ([dagger])   Right               5.0        Monosomy    Monosomy
                Left                4.6        Monosomy    Monosomy
 3 ([dagger])   Right#              9.1        Monosomy#   Monosomy#
   ([double       ([section])
   dagger])
                Left                2.2        Monosomy    Monosomy
 4 ([dagger])   Right               4.0        Monosomy    Monosomy
                Left            Site 1, 2.5#   Monosomy#   Monosomy#
                                ([section])
                                Site 2, NA     Not done    Not done
 5              Right               2.5        Monosomy    Monosomy
 6              Left                2.0        Monosomy    Monosomy
 7              Left#               3.7        Monosomy#   Monosomy#
                  ([section])
 8              Right               4.5        Disomy      Disomy
 9              Left                2.5        Monosomy    Disomy
10              Right               1.0        Monosomy    Disomy
11              Right               3.2        Disomy      Disomy
12              Left            Site 1, 3.5    Disomy      Disomy
13              Left                4.5        Disomy      Disomy
14              Left                3.5        Disomy      Disomy
15              NA                  NA         Disomy      Disomy
16              Right               1.9        Disomy      Disomy
17              Right               4.0        Disomy      Disomy
18              Right               4.7        Trisomy     Trisomy
19              Left                NA         Not done    Not done
20              Left                3.2        Not done    Not done
21              Left                1.3        Not done    Not done
22              Right               7.0        Not done    Not done

Case                Side         Size, cm             Karyotype

 1 ([dagger])   Right              12.2       43,X,-Y,-1,i(9)(p10),-21;
                                                [9],/47,XY,+7[2]/46,
                                                XY[9]
                Left                5.0       Not sampled
 2 ([dagger])   Right               5.0       43,X,-Y,-1,-14[6]/45,X,-Y
                                                [4]/46,XY[8]/NCA[2]
                Left                4.6       44,X,-Y,-1[20]
 3 ([dagger])   Right               9.1       47,XY,+18[3]/46,XY[12]/
   ([double       ([section])                   NCA[5]#
   dagger])
                Left                2.2       Not sampled
 4 ([dagger])   Right               4.0       Not sampled
                Left            Site 1, 2.5   46,XY[18]/NCA[2]#
                                ([section])
                                Site 2, NA    45,X,-Y[6]/46,XY[9]/NCA
                                                [5]
 5              Right               2.5       44,X,-X,-1[15]
 6              Left                2.0       44,XX,-1,-14[20]
 7              Left                3.7       46,XY[20]#
                  ([section])
 8              Right               4.5       46,XX[19]/NCA[1]
 9              Left                2.5       45,XX,der(1;13)(q10;q10),
                                                der(5)t(5;?21)(q11.2;
                                                ?p13),der(8)t(8;17)
                                                (q21.2;q23,der(17)
                                                t(5;17)(q33;q23),
                                                der(21)t(5;21)(q11.2;
                                                p13)t(5;8)(q33;q21.1),
                                                t(15;19)(q15;p13.1)
                                                [15]/45,idem,t(4;6)(p1
                                                ...6;q21)[2]/46,XX[3]
10              Right               1.0       46,XX,der(1;5)(q10;p10),
                                                +5[15]/46,XX[5]
11              Right               3.2       46,XY[20]
12              Left            Site 1, 3.5   46,XY[6]/NCA[1]
13              Left                4.5       45,X,-Y[3]/46,X,-Y,+7[2]/
                                                46,X,-Y,+14[1]/46,
                                                XY[14]
14              Left                3.5       46,XX[20]
15              NA                  NA        46,XY[20]
16              Right               1.9       46,XY[20]
17              Right               4.0       Not sampled
18              Right               4.7       No growth
19              Left                NA        45,X-Y[4]/46,XY[15]/47,
                                                XY,+X[1]
20              Left                3.2       46,XX,t(11;19)(p15;q12.2)
                                                [13]/46,XX[4]/NCA[3]
21              Left                1.3       46,XX[15]/NCA[5]
22              Right               7.0       46,XY[14]/45,X,-Y[3]/
                                                NCA[3]

* NCA indicates nonclonal abnormality; NA, not available.

([dagger]) Bilateral involvement.

([double dagger]) Renal oncocytosis.

([section]) Discordance between FISH analysis and conventional
cytogeneteic results, shown in bold.

Note: Discordance between FISH analysis and conventional cytogenetic
results are indicated with #.