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Clear cell sarcoma of the kidney: A genetical entity?
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Abstracts CLEAR CELL SARCOMA OF THE KIDNEY: A GENETICAL ENTITY? S. SZ0CS1 A. KOVACS2 and G. KOVACS2 PATHOLOGISCHESINSTITUT,TECHNISCHEUNIVERSIT.~,T (KLINIKUMRECHTSDERISAR)1 MUNCHEN,GERMANY ICRFCANCERGENETICLABORATORY'LONDON, , U.K.2
Recurring chromosomal alterations arc characteristic of human
AI0
HIGHLY REPETITIVE CHROMOSOME CHANGES ASSOCIATED WITH SKIN TUMOR DEVELOPM E N T IN T R A N S G E N I C M I C E T R E A T E D W I T H TPA.
Bisharah L. Libbus" John E. French b, Judson Spalding, Laura Hansen b, Raymond R. TicK, and Raymond W. Tennant b. ~ e n e t l c Research, Inc., RTP, NC 27709; bNational Institute of Environmental Health Sciences, RTP, NC 27709. CIntegrated Laboratory Systems, RTP, NC 27709.
diffcrent from those found in Wilms tumor and RCC.
Cytogenetic changes associated with skin tumorigenesis were examined in transgenic mice carrying an a c t i v a t e d v-Ha-ras gene. Following multiple applications of tumor promotor TPA twice weekly for 11 weeks, papillomas appear and some later convert to skin tumors. A number of such tumors were excised, cultured and analyzed for the presence of numerical or structural chromosome abnormalities. In all eight tumors analyzed, extra copies of chromosome 10 were present in 90-100% of the cells karyotyped. Trisomy 6 was observed in 6 of 8 tumors studied. On the other hand, monosmoy 13 and 14 was present in 7 and 6 tumors respectively. Structural rearrangements were infrequently observed and no indications of gene amplification was noted. Cultured oells were highly tumorigenic when transplanted to 9-11 day old syngeneic recipients, resulting in large ulcerating tumors within two weeks. Western analysis of transplanted tumors indicated high expression levels of the transgene. These results indicate that tumor cells initiated by insertion of this v-Ha-ras transgene followed by chemical promotion involve specific chromosome changes which Ray be important in identifying other oncogenes and/or tumor suppressor genes involved in skin tumorigenesis. Supported by NIEHS contract N01-ES-15321.
All
A12
tumors and they play an important role in the diagnosis and classification in hematologic neoplasms. Although much less cytogcncnc data exists in human solid tumors, malignancies of the kidney are gcnctically well charactcrised such as 3p rearrangement in renal ccll carcinoma (RCC) or the alteration of 11p in Wilms tumor. Moreover, thc cytogenctic and moleculargcnctic analysis of RCC clearly showed the different entity of histomorphologic manifestations (e. g.papillary and non papillary RCC). Clear cell sarcoma of the kidney (CCSK) is a rare form of human rcnal tumor dcrived from mcscnchymal cells and not widely recognised. We analysed a (CCSK), diagnosed in a 21 year old male, by means of cytogenetic and molecular genetic methods. 25 mctaphascs were karyotyped. A balanced translocation t(4;13) (p14; q32) was found in each tumor cell, while the normal kidney cells and the peripheral blood cells had a normal karyotype. Loss of heterozygosity at 11 p, or rearrangement of chromosome 3p region, was not detected. No gross alterations in the structure and expression of the Wilms tumor gene were found. Our findings suggcst that CCSK might have molecular genetic alterations
LOSS OF HETEROZYGOSITY AFFECTING CHROMOSOME 1 IN R A T M A M M A R Y TUMORS ASSOCIATION WITH CHROMOSOME OVERREPRESENTATION AND ONCOGENE A C T I V A T I O N . C.M. Aldaz, L.S. Gollahon and A. Chen. University of Texas M.D. Anderson Cancer Center, Science Park - Research Division, Smithville, Texas. The rat m n m m a r y carcinogenesis model is one of the most widely used systems for the study of msrnmqry cancer. Very recently we have shown the nonrandom nature of abnormalities affecting rat chromosome I during mamrn~'y tumor progression. W e observed and association between chromosome 1 overrepresentation (i.e.direct duplications or trisomies), progressive loss of differentiationof the tumors and amplification of the mutated H a - m s oncogene. B y analyzing tumors induced in F1 hybrid rats w e n o w have also observed loss of h e t e r o z y g o s i t y a t d i f f e r e n t loci w i t h i n r a t chromosome 1. W e are currently studying the association of t h e L O H w i t h t h e previous findings affecting t h e s a m e c h r o m o s o m e . T h e m a i n q u e s t i o n is w h e t h e r t h e o b s e r v e d L O H is t h e c o n s e q u e n c e of r e c o m b i n a t i o n or n o n d i s j u n c t i o n e v e n t s l e a d i n g to t h e selection of t h e c h r o m o s o m e b e a r i n g t h e m u t a t e d oncogene or if i n a d d i t i o n r a t c h r o m o s o m e I m a y be the locale of a putative tumor suppressor gene. T h e s t r o n g s i m i l a r i t i e s w i t h p r e v i o u s f i n d i n g s in m o u s e s k i n c a r c i n o g e n e s i s s u g g e s t t h a t t h e s e m a y be relevant mechanisms behind the very common t r i s o m i e s a n d d u p l i c a t i o n s o b s e r v e d in solid t u m o r s . S u p p o r t e d by NI-II g r a n t CA 48922
A BREAKPOINT ON CHROMOSOME 6q IS INVOLVED IN CELL IMMORTALIZATION P. Patsalis', H. Ozer=, and A. Henderson'. 'Hunter CollegeCity University of New York, New York ; 2 UMDNJ-New Jersey Medical School, Newark, NJ. Transformation of human diploid fibroblasts (HF) with SV40 can result in extension of life span beyond the normal limit of senescence and in a minority of cases, immortalization. This stu0y used comparison of matched parental (diploid) and immortalized cell lines to determine if any single factor could be relate¢l to the immortalization phenomena. The integration site of SV40 was shown to be at chromosome 5o,21 by cytologic hybridization. A comparison of mortal and immortal cells showed no alterations involving the integrated SV40 genome per se. A survey of random tranalocations and other anomalies occurring in the immortalized lines was allo made. Some of these are regions known to contain ormegenas and transforming proteins. These were not rearranged. Karyotypic analysis of matctled cell lines, however, identified a specific chromosomal break~int (6o,21) in immortalizedcells that was not preaent in the pimlntal line. Southem blot analysis conformed that xquencml on the distal portion of 6(I are lost in i,vnettalic~d ~ DNA which flanks the braakpoint has been identHled. U ~ the deleted DNA from the bre~point,a YAC library was screened and two positive donas were [sol=ted. Further analysis of these clones will attempt to determine the genes in the breakpoint region and study their s i g ~ in immortalization. We propose that deletion of specific sequences due to breakage at fragile sites of chromosome 6q represent one of the mutational events responsible for immortalization of SV40 transformed HF.
A9
Abstracts CLEAR CELL SARCOMA OF THE KIDNEY: A GENETICAL ENTITY? S. SZ0CS1 A. KOVACS2 and G. KOVACS2 PATHOLOGISCHESINSTITUT,TECHNISCHEUNIVERSIT.~,T (KLINIKUMRECHTSDERISAR)1 MUNCHEN,GERMANY ICRFCANCERGENETICLABORATORY'LONDON, , U.K.2
Recurring chromosomal alterations arc characteristic of human
AI0
HIGHLY REPETITIVE CHROMOSOME CHANGES ASSOCIATED WITH SKIN TUMOR DEVELOPM E N T IN T R A N S G E N I C M I C E T R E A T E D W I T H TPA.
Bisharah L. Libbus" John E. French b, Judson Spalding, Laura Hansen b, Raymond R. TicK, and Raymond W. Tennant b. ~ e n e t l c Research, Inc., RTP, NC 27709; bNational Institute of Environmental Health Sciences, RTP, NC 27709. CIntegrated Laboratory Systems, RTP, NC 27709.
diffcrent from those found in Wilms tumor and RCC.
Cytogenetic changes associated with skin tumorigenesis were examined in transgenic mice carrying an a c t i v a t e d v-Ha-ras gene. Following multiple applications of tumor promotor TPA twice weekly for 11 weeks, papillomas appear and some later convert to skin tumors. A number of such tumors were excised, cultured and analyzed for the presence of numerical or structural chromosome abnormalities. In all eight tumors analyzed, extra copies of chromosome 10 were present in 90-100% of the cells karyotyped. Trisomy 6 was observed in 6 of 8 tumors studied. On the other hand, monosmoy 13 and 14 was present in 7 and 6 tumors respectively. Structural rearrangements were infrequently observed and no indications of gene amplification was noted. Cultured oells were highly tumorigenic when transplanted to 9-11 day old syngeneic recipients, resulting in large ulcerating tumors within two weeks. Western analysis of transplanted tumors indicated high expression levels of the transgene. These results indicate that tumor cells initiated by insertion of this v-Ha-ras transgene followed by chemical promotion involve specific chromosome changes which Ray be important in identifying other oncogenes and/or tumor suppressor genes involved in skin tumorigenesis. Supported by NIEHS contract N01-ES-15321.
All
A12
tumors and they play an important role in the diagnosis and classification in hematologic neoplasms. Although much less cytogcncnc data exists in human solid tumors, malignancies of the kidney are gcnctically well charactcrised such as 3p rearrangement in renal ccll carcinoma (RCC) or the alteration of 11p in Wilms tumor. Moreover, thc cytogenctic and moleculargcnctic analysis of RCC clearly showed the different entity of histomorphologic manifestations (e. g.papillary and non papillary RCC). Clear cell sarcoma of the kidney (CCSK) is a rare form of human rcnal tumor dcrived from mcscnchymal cells and not widely recognised. We analysed a (CCSK), diagnosed in a 21 year old male, by means of cytogenetic and molecular genetic methods. 25 mctaphascs were karyotyped. A balanced translocation t(4;13) (p14; q32) was found in each tumor cell, while the normal kidney cells and the peripheral blood cells had a normal karyotype. Loss of heterozygosity at 11 p, or rearrangement of chromosome 3p region, was not detected. No gross alterations in the structure and expression of the Wilms tumor gene were found. Our findings suggcst that CCSK might have molecular genetic alterations
LOSS OF HETEROZYGOSITY AFFECTING CHROMOSOME 1 IN R A T M A M M A R Y TUMORS ASSOCIATION WITH CHROMOSOME OVERREPRESENTATION AND ONCOGENE A C T I V A T I O N . C.M. Aldaz, L.S. Gollahon and A. Chen. University of Texas M.D. Anderson Cancer Center, Science Park - Research Division, Smithville, Texas. The rat m n m m a r y carcinogenesis model is one of the most widely used systems for the study of msrnmqry cancer. Very recently we have shown the nonrandom nature of abnormalities affecting rat chromosome I during mamrn~'y tumor progression. W e observed and association between chromosome 1 overrepresentation (i.e.direct duplications or trisomies), progressive loss of differentiationof the tumors and amplification of the mutated H a - m s oncogene. B y analyzing tumors induced in F1 hybrid rats w e n o w have also observed loss of h e t e r o z y g o s i t y a t d i f f e r e n t loci w i t h i n r a t chromosome 1. W e are currently studying the association of t h e L O H w i t h t h e previous findings affecting t h e s a m e c h r o m o s o m e . T h e m a i n q u e s t i o n is w h e t h e r t h e o b s e r v e d L O H is t h e c o n s e q u e n c e of r e c o m b i n a t i o n or n o n d i s j u n c t i o n e v e n t s l e a d i n g to t h e selection of t h e c h r o m o s o m e b e a r i n g t h e m u t a t e d oncogene or if i n a d d i t i o n r a t c h r o m o s o m e I m a y be the locale of a putative tumor suppressor gene. T h e s t r o n g s i m i l a r i t i e s w i t h p r e v i o u s f i n d i n g s in m o u s e s k i n c a r c i n o g e n e s i s s u g g e s t t h a t t h e s e m a y be relevant mechanisms behind the very common t r i s o m i e s a n d d u p l i c a t i o n s o b s e r v e d in solid t u m o r s . S u p p o r t e d by NI-II g r a n t CA 48922
A BREAKPOINT ON CHROMOSOME 6q IS INVOLVED IN CELL IMMORTALIZATION P. Patsalis', H. Ozer=, and A. Henderson'. 'Hunter CollegeCity University of New York, New York ; 2 UMDNJ-New Jersey Medical School, Newark, NJ. Transformation of human diploid fibroblasts (HF) with SV40 can result in extension of life span beyond the normal limit of senescence and in a minority of cases, immortalization. This stu0y used comparison of matched parental (diploid) and immortalized cell lines to determine if any single factor could be relate¢l to the immortalization phenomena. The integration site of SV40 was shown to be at chromosome 5o,21 by cytologic hybridization. A comparison of mortal and immortal cells showed no alterations involving the integrated SV40 genome per se. A survey of random tranalocations and other anomalies occurring in the immortalized lines was allo made. Some of these are regions known to contain ormegenas and transforming proteins. These were not rearranged. Karyotypic analysis of matctled cell lines, however, identified a specific chromosomal break~int (6o,21) in immortalizedcells that was not preaent in the pimlntal line. Southem blot analysis conformed that xquencml on the distal portion of 6(I are lost in i,vnettalic~d ~ DNA which flanks the braakpoint has been identHled. U ~ the deleted DNA from the bre~point,a YAC library was screened and two positive donas were [sol=ted. Further analysis of these clones will attempt to determine the genes in the breakpoint region and study their s i g ~ in immortalization. We propose that deletion of specific sequences due to breakage at fragile sites of chromosome 6q represent one of the mutational events responsible for immortalization of SV40 transformed HF.