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Trisomy 20 in a papillary urothelial carcinoma of the ureter
ELSEVIER
Trisomy 20 in a Papillary Urothelial Carcinoma of the Ureter Paola Grammatico, Antonietta Lombardo, Massimo Governatori, Cristina Mordenti, Monica Poscente, and Giuseppe Del Porto
ABSTRACT: To contribute to the knowledge on tumorigenesis and the evolution of urothelial carcinoma of the ureter, we analyzed the clinical, histological, and cytogenetic aspects of a case. Primary cell cultures obtained from tumor specimens showed a trisomy of chromosome 20 where the c-src protooncogene, already described in literature as having an important role in the etiology and progression of some tumors, is located. In our case trisomy 20 is the only present marker and for this reason we think that it could play a role in the tumorigenesis of the urothelial carcinoma of the ureter.
INTRODUCTION
Urothelial carcinoma of the ureter is a rare tumor that presents a wide clinical heterogeneity, with particular regard to evolution. Up to date grading and staging are the most useful parameters for prognosis, but in some cases they are considered insufficient. To explain the cases with an atypical evolution, during the last years some authors have found further characteristics that could allow them to obtain a subclassification under genetic aspects. Berger et al. [1], in a review regarding the cytogenetic studies of urinary tumors, described nine cases that showed a trisomy of chromosome 20. In particular it showed one transitional cell carcinoma, six renal cell carcinomas, and two transitional cell carcinomas of the bladder. In all these cases the trisomy never appeared as the only present aberration and, for this reason, the authors suggested that it could be considered a secondary change. Trisomy 20 was also described, in addition to other chromosomal abnormalities, by other authors [2-7] who analyzed various tumors of the urinary tract. In 1987 Smeets et al. [8] described 13 cases of primary transitional cell carcinomas of the bladder. In two lesions obtained from a single site, at a distance of six months, a trisomy of chromosome 20 was found as the only present aberration in a patient.
From the Experimental Medicine and Pathology Department, Section of Medical Genetics (P. G., A. L., C. M., M. P., G. D. P.), University "La Sapienza," Rome, Italy, and Malpighi Urological Division (M. G.), Osp. S. Camille, Rome, Italy. Address reprint requests to: Prof. Giuseppe DeI Porto, Cattedra di Genetiea Medica, Universitgl "La Sapienza, " c/o Osp. L. Spallanzani, Via Portuense n. 292, 00149, Roma Italia. Received August 7, 1995; accepted February 23, 1996. Cancer Genet Cytogenet 90:132-134 (1996) © Elsevier Science Inc., 1996 655 Avenue of the Americas, N e w York, NY 10010
To contribute to the knowledge of the etiology and progression of this neoplasm and to understand the role of some particular chromosomal involvement, we performed a cytogenetic study on a case of urothelial carcinoma of the ureter, clinically and histologically defined, and compared our data with the results already present in literature.
CASE REPORT
The patient (M.A.) was a 55-year-old male. He showed a 1 cm tumor, defined as a microinvasive papillary urothelial carcinoma, at 2.7 cm from the right distal margin of the ureter's surgery resection. After surgical treatment the patient underwent a clinical and instrumental follow-up to evaluate the possible development of the disease. About six years later he appears to be free from any relapse.
MATERIALS AND METHODS
Cytogenetic analysis was conducted on peripheral blood lymphocytes to exclude the presence of a possible constitutional aberration. The primary cell cultures from the tumor specimen were obtained after mechanical dissection of the neoplastic fragment using RPMI 1640 mixed with fetal bovine serum (15%), L-glutamine (1%), and penicillin-streptomycin (50 Ui/ml-50 txg/ml). To characterize the origin of the cells, we performed an immunocytochemical test using the URO1 antibody (Clone J143) that detects an antigen expressed in the basement membrane of the glomerulus and urothelium, and recognizes bladder tumors and other epithelial neoplasms. The metaphases were obtained after an overnight
0165-4608/96/$15.00 PII S0165-4608(96)00073-8
Trisomy 20 in Urothelial Ca:rcinomas
133
colchicine treatment followed by a hypotonic shock with KC1 (0.075 M) and fixation using a 3:1 methanol-acetic acid solution. The karyotype was performed on 66 cells using GTG- and CBG-banding techniques. RESULTS
Histological study showed a microinvasive papillary urothelial carcinoma G2, pT1/T2 NO M0 (Fig. 1 a,b). Constitutional karyotype obtained from peripheral blood lym~ phocytes was: 46,XY. Immunocytochemical analyses conducted on a sample from the primary cell cultures demonstrated a positive result for URO1 confirming tile origin of the cells. Chromosome study conducted on three different primary cell cultures shows a trisomy of chromosome 20 (Fig. 2) in 44 of the 66 anal~yzed cells. DISCUSSION
Cytogenetic results obtained during the last few years on different solid tumors have contributed to the identificaFigure 1 (a, b) Histological appearance of the papillary urothelial carcinoma. Hemotoxylin/eosin. i
a
b
Figure 2 Trisomy20, GTG-banded metaphase.
tion of the chromosomal regions involved in the tumorigenesis and in the progression of the neoplasms [9, 10]. Detection of the abnormalities that occur as single markers in more cases of the same tumor is important for the characterization of the chromosomal regions involved in the tumorigenesis other than in the evolution of the disease. In our case trisomy 20 is the only present aberration. For this reason, and because the same chromosomal aberration was the only present marker in a case of bladder cancer reported by Smeets et al. [8], and appeared to be very frequent in the literature regarding cytogenetic studies on tumors affecting urothelial cancer [1-7], we believe that this abnormality could be considered a primary change. In addition we also have to consider that our patient is in good health after six years. We think it's important to stress that Sakaguchi et al. [11] mapped the c-src on chromosome 20, a proto-oncogene that has often been found associated with human neoplasms. Fanning et al. [12] highlighted that a subtype of bladder tumors, the low-grade lesions, showed high levels of pp60csrc expression and suggested a possible role of the src proto-oncogene in urotheiial cell differentiation processes. In addition, Quintrell et al. [13] identified the HCK gene, that encodes a protein-tyrosin kinase that resembles the product of the proto-oncogene c-src. HCK is localized on 20q11-q12, which is often involved in some acute ]eukemias and myeloproliferative disorders. In conclusion, we suggest that trisomy 20 could play an important role in the etiology and pathogenesis of urothelial carcinoma of the ureter because of the overexpression of c-src and HCK sequences. In addition we think it is important to encourage the publication of further cases with trisomy 20 as the only present marker, well characterized for clinical follow-up concerns, to understand if it is possible to consider this marker as a positive prognostic parameter. If this hypothe-
134
P. G r a m m a t i c o et al.
sis is c o n f i r m e d in the future w e c o u l d use c y t o g e n e t i c s t u d y as a further i n d e x for a m o r e p r e c i s e prognosis.
7. Dal Cin P, Li FP, Prout GR, Huben RR Limon J, Ferti-Passantonopoulou A, Richie JP, Sandberg AA (1988): Involvement of chromosome 3 and 5 in renal cell carcinoma. Cancer Genet Cytogenet 35:41-46.
REFERENCES
8. Smeets W, Pauwels R, Laarakkers D, Debruyne F, Geraedts J (1987): Chromosomal analysis of bladder cancer. III. Nonrandom alterations. Cancer Genet Cytogenet 29:29-41.
1. Berger CS, Sandberg AA, Todd IAD, Pennington RD, Haddad FS, Hecht BK, Hecht F (1986): Chromosomes in kidney, ureter, and bladder cancer. Cancer Genet Cytogenet 23:1-24. 2. Atkin NB, Baker MC (1985): Cytogenetic study of ten carcinomas of the bladder: Involvement of chromosome 1 and 11. Cancer Ganet Cytogenet 15:253. 3. Meloni AM, Dobbs RM, Pontes JE, Sandberg AA (1993): Translocation (X;1) ill papillary renal cell carcinoma. A new cytogenetic subtype. Cancer Genet Cytogenet 65:1-6. 4. De long B, Oosterhuis IW, Idenburg V]S, Castedo SMM], Dam A, Mensink H]A [1988): Cytogenetics of 12 cases of renal adenocarcinoma. Cancer Genet Cytogenet 30:53-61. 5. Kovacs G, Szucs S, De Riese W, Baumgarte] H (1987): Specific chromosome aberration in human renal cell carcinoma. Int l Cancer 40:171-178. 6. Kovacs GM, Frish S (1989): Clonal chromosome abnormalities in tumor cells from patients with sporadic renal cell carcinomas. Cancer Res 49:651-659.
9. Meloni AM, Bridge J, Sandberg AA (1992): Reviews on chromosome studies in urological tumors. I. Renal Tumors. J Urol 148:253-265. 10. Sandberg AA, Berger CS (1994): Review of chromosome studies in urological tumors, lI. Cytogenetics and molecular genetics of bladder cancer. I Urol 151:545-560. 11. Sakaguchi AY, Naylor SL, Shows TB (1983): A sequence homologous to rous sarcoma Virus v-src is on human chromosome 20. Prog Nucleic Acid Res Mol Biol 29:279-283. 12. Fanning P, Bulovas K, Saini KS, Libertino JA, Joyce AD, Summerhayes IC (1992): Elevated expression of pp60c-src in low grade bladder carcinomas. Cancer Res 52:1457-1462. 13. Quintrell N, Lebo R, Varmus H, Bishop M, Pettenati M, Le Beau M, Diaz MO, Rowley J (1987): Identification of a human gene (HCK) that encodes a protein-tyrosine kinase and is expressed in hemopoietic cells. Mol Cell Biol 2267-2275.
Trisomy 20 in a Papillary Urothelial Carcinoma of the Ureter Paola Grammatico, Antonietta Lombardo, Massimo Governatori, Cristina Mordenti, Monica Poscente, and Giuseppe Del Porto
ABSTRACT: To contribute to the knowledge on tumorigenesis and the evolution of urothelial carcinoma of the ureter, we analyzed the clinical, histological, and cytogenetic aspects of a case. Primary cell cultures obtained from tumor specimens showed a trisomy of chromosome 20 where the c-src protooncogene, already described in literature as having an important role in the etiology and progression of some tumors, is located. In our case trisomy 20 is the only present marker and for this reason we think that it could play a role in the tumorigenesis of the urothelial carcinoma of the ureter.
INTRODUCTION
Urothelial carcinoma of the ureter is a rare tumor that presents a wide clinical heterogeneity, with particular regard to evolution. Up to date grading and staging are the most useful parameters for prognosis, but in some cases they are considered insufficient. To explain the cases with an atypical evolution, during the last years some authors have found further characteristics that could allow them to obtain a subclassification under genetic aspects. Berger et al. [1], in a review regarding the cytogenetic studies of urinary tumors, described nine cases that showed a trisomy of chromosome 20. In particular it showed one transitional cell carcinoma, six renal cell carcinomas, and two transitional cell carcinomas of the bladder. In all these cases the trisomy never appeared as the only present aberration and, for this reason, the authors suggested that it could be considered a secondary change. Trisomy 20 was also described, in addition to other chromosomal abnormalities, by other authors [2-7] who analyzed various tumors of the urinary tract. In 1987 Smeets et al. [8] described 13 cases of primary transitional cell carcinomas of the bladder. In two lesions obtained from a single site, at a distance of six months, a trisomy of chromosome 20 was found as the only present aberration in a patient.
From the Experimental Medicine and Pathology Department, Section of Medical Genetics (P. G., A. L., C. M., M. P., G. D. P.), University "La Sapienza," Rome, Italy, and Malpighi Urological Division (M. G.), Osp. S. Camille, Rome, Italy. Address reprint requests to: Prof. Giuseppe DeI Porto, Cattedra di Genetiea Medica, Universitgl "La Sapienza, " c/o Osp. L. Spallanzani, Via Portuense n. 292, 00149, Roma Italia. Received August 7, 1995; accepted February 23, 1996. Cancer Genet Cytogenet 90:132-134 (1996) © Elsevier Science Inc., 1996 655 Avenue of the Americas, N e w York, NY 10010
To contribute to the knowledge of the etiology and progression of this neoplasm and to understand the role of some particular chromosomal involvement, we performed a cytogenetic study on a case of urothelial carcinoma of the ureter, clinically and histologically defined, and compared our data with the results already present in literature.
CASE REPORT
The patient (M.A.) was a 55-year-old male. He showed a 1 cm tumor, defined as a microinvasive papillary urothelial carcinoma, at 2.7 cm from the right distal margin of the ureter's surgery resection. After surgical treatment the patient underwent a clinical and instrumental follow-up to evaluate the possible development of the disease. About six years later he appears to be free from any relapse.
MATERIALS AND METHODS
Cytogenetic analysis was conducted on peripheral blood lymphocytes to exclude the presence of a possible constitutional aberration. The primary cell cultures from the tumor specimen were obtained after mechanical dissection of the neoplastic fragment using RPMI 1640 mixed with fetal bovine serum (15%), L-glutamine (1%), and penicillin-streptomycin (50 Ui/ml-50 txg/ml). To characterize the origin of the cells, we performed an immunocytochemical test using the URO1 antibody (Clone J143) that detects an antigen expressed in the basement membrane of the glomerulus and urothelium, and recognizes bladder tumors and other epithelial neoplasms. The metaphases were obtained after an overnight
0165-4608/96/$15.00 PII S0165-4608(96)00073-8
Trisomy 20 in Urothelial Ca:rcinomas
133
colchicine treatment followed by a hypotonic shock with KC1 (0.075 M) and fixation using a 3:1 methanol-acetic acid solution. The karyotype was performed on 66 cells using GTG- and CBG-banding techniques. RESULTS
Histological study showed a microinvasive papillary urothelial carcinoma G2, pT1/T2 NO M0 (Fig. 1 a,b). Constitutional karyotype obtained from peripheral blood lym~ phocytes was: 46,XY. Immunocytochemical analyses conducted on a sample from the primary cell cultures demonstrated a positive result for URO1 confirming tile origin of the cells. Chromosome study conducted on three different primary cell cultures shows a trisomy of chromosome 20 (Fig. 2) in 44 of the 66 anal~yzed cells. DISCUSSION
Cytogenetic results obtained during the last few years on different solid tumors have contributed to the identificaFigure 1 (a, b) Histological appearance of the papillary urothelial carcinoma. Hemotoxylin/eosin. i
a
b
Figure 2 Trisomy20, GTG-banded metaphase.
tion of the chromosomal regions involved in the tumorigenesis and in the progression of the neoplasms [9, 10]. Detection of the abnormalities that occur as single markers in more cases of the same tumor is important for the characterization of the chromosomal regions involved in the tumorigenesis other than in the evolution of the disease. In our case trisomy 20 is the only present aberration. For this reason, and because the same chromosomal aberration was the only present marker in a case of bladder cancer reported by Smeets et al. [8], and appeared to be very frequent in the literature regarding cytogenetic studies on tumors affecting urothelial cancer [1-7], we believe that this abnormality could be considered a primary change. In addition we also have to consider that our patient is in good health after six years. We think it's important to stress that Sakaguchi et al. [11] mapped the c-src on chromosome 20, a proto-oncogene that has often been found associated with human neoplasms. Fanning et al. [12] highlighted that a subtype of bladder tumors, the low-grade lesions, showed high levels of pp60csrc expression and suggested a possible role of the src proto-oncogene in urotheiial cell differentiation processes. In addition, Quintrell et al. [13] identified the HCK gene, that encodes a protein-tyrosin kinase that resembles the product of the proto-oncogene c-src. HCK is localized on 20q11-q12, which is often involved in some acute ]eukemias and myeloproliferative disorders. In conclusion, we suggest that trisomy 20 could play an important role in the etiology and pathogenesis of urothelial carcinoma of the ureter because of the overexpression of c-src and HCK sequences. In addition we think it is important to encourage the publication of further cases with trisomy 20 as the only present marker, well characterized for clinical follow-up concerns, to understand if it is possible to consider this marker as a positive prognostic parameter. If this hypothe-
134
P. G r a m m a t i c o et al.
sis is c o n f i r m e d in the future w e c o u l d use c y t o g e n e t i c s t u d y as a further i n d e x for a m o r e p r e c i s e prognosis.
7. Dal Cin P, Li FP, Prout GR, Huben RR Limon J, Ferti-Passantonopoulou A, Richie JP, Sandberg AA (1988): Involvement of chromosome 3 and 5 in renal cell carcinoma. Cancer Genet Cytogenet 35:41-46.
REFERENCES
8. Smeets W, Pauwels R, Laarakkers D, Debruyne F, Geraedts J (1987): Chromosomal analysis of bladder cancer. III. Nonrandom alterations. Cancer Genet Cytogenet 29:29-41.
1. Berger CS, Sandberg AA, Todd IAD, Pennington RD, Haddad FS, Hecht BK, Hecht F (1986): Chromosomes in kidney, ureter, and bladder cancer. Cancer Genet Cytogenet 23:1-24. 2. Atkin NB, Baker MC (1985): Cytogenetic study of ten carcinomas of the bladder: Involvement of chromosome 1 and 11. Cancer Ganet Cytogenet 15:253. 3. Meloni AM, Dobbs RM, Pontes JE, Sandberg AA (1993): Translocation (X;1) ill papillary renal cell carcinoma. A new cytogenetic subtype. Cancer Genet Cytogenet 65:1-6. 4. De long B, Oosterhuis IW, Idenburg V]S, Castedo SMM], Dam A, Mensink H]A [1988): Cytogenetics of 12 cases of renal adenocarcinoma. Cancer Genet Cytogenet 30:53-61. 5. Kovacs G, Szucs S, De Riese W, Baumgarte] H (1987): Specific chromosome aberration in human renal cell carcinoma. Int l Cancer 40:171-178. 6. Kovacs GM, Frish S (1989): Clonal chromosome abnormalities in tumor cells from patients with sporadic renal cell carcinomas. Cancer Res 49:651-659.
9. Meloni AM, Bridge J, Sandberg AA (1992): Reviews on chromosome studies in urological tumors. I. Renal Tumors. J Urol 148:253-265. 10. Sandberg AA, Berger CS (1994): Review of chromosome studies in urological tumors, lI. Cytogenetics and molecular genetics of bladder cancer. I Urol 151:545-560. 11. Sakaguchi AY, Naylor SL, Shows TB (1983): A sequence homologous to rous sarcoma Virus v-src is on human chromosome 20. Prog Nucleic Acid Res Mol Biol 29:279-283. 12. Fanning P, Bulovas K, Saini KS, Libertino JA, Joyce AD, Summerhayes IC (1992): Elevated expression of pp60c-src in low grade bladder carcinomas. Cancer Res 52:1457-1462. 13. Quintrell N, Lebo R, Varmus H, Bishop M, Pettenati M, Le Beau M, Diaz MO, Rowley J (1987): Identification of a human gene (HCK) that encodes a protein-tyrosine kinase and is expressed in hemopoietic cells. Mol Cell Biol 2267-2275.