Chromosome 9 Allelic Losses and Microsatellite Alterations in Human Bladder Tumors'

[CANCER RESEARCH54, 2848-2851, June 1, 19941 Advances in Brief

Irene Orlow, Pilar Lianes, Louis Lacombe, Guido Dalbagni, Victor E. Reuter, and Carlos Cordon-Cardo2

Department of Pathology [I. 0., P. L., V. E. R., C. C-C.] and Urology Service [L L., G. D.], Memorial Sloan-Kettering Cancer Center, New York New York 10021

Abstract the inactivation of a tumor suppressor gene (12â€”14).The inherited basis of these neoplasia appears to be due to an inactivation of DNA Chromosome 9 allelic losses have been reported as a frequent and early damage recognition/repair pathways. However, even in hereditary eventoccurringinbladdercancer.It hasbeenpostulatedthata candidate nonpolyposis colorectal cancers, multiple hits are necessary for the tumor suppressor gene may reside on this chromosome, alterations of which may lead to the development of a subset of superficial bladder transformed cells to acquire the malignant phenotype. The objectives tumors.Morerecently,theinvolvementoftwodifferentregionsharboring of this study were to better define the deleted regions of the chromo suppressor loci, one on each of both chromosome 9 arms, has been some 9 in bladder tumors, to delineate the possible role of the loss of proposed. We undertook the present study with the objectives of better heterozygosity (LOH3) in 9p and 9q arms, and to evaluate the exis defining the deleted regions of chromosome 9 in bladder tumors, as well tence and frequency of microsatellite alterations in urothelial neopla as evaluatingthe frequencyof microsateffitealterationsaffectingcertain sias. We examined genetic alterations occurring on chromosome 9 loci on this chromosome in urothelial neoplasia. Seventy-threeprimary using a well-characterized cohort of patients affected with superficial bladdertumorswereanalyzedusinga setofhighlypolymorphicmarkers, and muscle invasive bladder tumors. We centered our study on the andresultswerecorrelatedwithpathologicalparametersassociatedwith analyses of RFLP, VNTR, and microsatellite markers assigned to both poor Clinical outcome. We observed that, overall, 77% of the tumors arms of chromosome 9. studied showed either loss of heterozygosity for one or more chromosome 9 markers and/or microsteffite abnormalities at chromosome9 loci. De tailed analyses showed that two regions, one on 9p at the interferon Materials and Methods cluster, and the other on 9q associated with the q34.1â€”2bands, had the Tissues. Seventy-three primary bladder tumors were obtained from rad highest frequencies of allelic losses. Furthermore, T@lesions appeared to ical cystectomy samples (n = 64) and transurethral resection specimens present mainly with 9q abnormalities, while T1 tumors displayed a mix (n 9). Tissues were embedded in tissue TEK OCI' cryopreservative tare of aberrant9p and 9q genotypes.Theseobservationsindicatethat (Miles, Inc., Elkhart, IN) and stored frozen at â€”70Â°C.Normaltissue was loss of heterozygosity of 9p may be associated with the development of obtained from the same bladder in an area free of tumor or, in 5 cases, from superficial tumors with a more aggressive biological behavior or, alter other normal tissue (i.e., peripheral blood or prostate). Five-p@msections natively, they may be related to early disease progression. In addition, were used for hematoxylin and eosin staining. The sections were examined microsatellite alterations were documented in over 40% of amplified by a pathologist (V. E. R.) to confirm the presence of tumor, evaluation of cases. Taken together, these data suggest that two different tumor sup tissue morphology, and pathology staging. The 73 cases selected showed at pressor gene loci on chromosome 9 are involved as tumorigenic events in least 50% of tumor cells on the sample and were included in the study. All bladder cancer and that chromosome 9 microsatellite alterations are specimens were graded by WHO classification and staged according to the frequent events occurring in urothelial neoplasia. ThM pathological staging system. Introduction DNAIsolationandSouthernBlotting.Twentyto30consecutive30-g.irn thick sections were cut from each tissue block. The DNA was isolated, Cytogenetic and molecular genetic analyses of bladder cancer have digested, and transferred to nylon membrane as described previously (2, 3). identified abnormalities in a number of chromosomes which appear to The DNA was fixed to the membrane using a UV linker (UV Stratalink 1800; be involved in the development and progression of these tumors Stratagene, La Jolla, CA). (1â€”4).Loss of heterozygosity of chromosome 9 is a frequent mutation Probes. The following probes with chromosome map positions, loci, and restriction enzymes were obtained from the ATCC (Rockville, MD) and were occurring in both superficial and muscle invasive lesions. Allelic used for RFLP analysis in this study: p08782 (9q34â€”qter; Assgl; HindIII); losses in 9p (5â€”7)and 9q (3, 8, 9) have been reported, estimating the pabl K2 (9q34; v-nb!; TaqI); pMCOA12 (9q; D9S28; MspI); p04580 (9q34.3; area of deletions between 9p22 and 9q34.1. The involvement of two D9SJO; TaqI) VNTR; pEFD126.3 (9q34; D9S7; TaqI) VN'FR; DR6 different regions containing suppressor loci in bladder tumors has (9p21 â€”ter;D9S3; Hind!!!) (provided by Jane Fountain, Massachusetts Insti been recently proposed (10). In addition, somatic instability at mic tate of Technology-Center for Cancer Research, Cambridge, MA); pINIF A3 rosatellite repeats, detected in low stage bladder tumors, has been (9p22; interferon a-2; MspI) (provided by Dr.P.M. Pitha Rove, The John reported as an early alteration associated with tumorigenic events in Hopkins University-Oncology Center, Baltimore, MD). such neoplasias (11). The microsatellite instability observed in the Primers. The following primers were used for the present study: IFNA hereditary nonpolyposis colorectal cancers (Lynch syndrome) sug cluster (5'-TGCGCGTFAAGTTAA11XIG1T-3' and 5'-AGTGGGGGlTf C- gested that these tumors may arise through a mechanism different than CACCITAC-3', provided by Dr. Manuel Diaz, Loyola University School of Medicine, Chicago, IL); D9S54 (ATCC); ABL 1 (9q34.1) (ATCC); and Ass (9q34.1) (ATCC). Received 2/28/94; accepted 4/21/94. Thecostsof publicationofthisarticleweredefrayedinpartby thepaymentofpage Hybridization, PCR Amplification, and Autoradiography. The probes charges. This article must therefore be hereby marked advertisement in accordance with were labeled to a high specific activity with [a-32P]dCTP by the random 18 U.S.C. Section 1734 solely to indicate this fact. oligolabeling technique using the Primed DNA Labeling kit (Boehringer I This work was in part supported by NCI Grant CA-47538 (to C. C. C.). P. L is the Mannheim, Indianapolis, IN), and membranes were pretreated and hybridized recipient of a Postdoctoral Fellowship from the Spanish Government (P155 93/5052) and an NCI/EORTC Award. L L is the recipient of a Fellowship Award from the Kidney

Foundation of Canada. 3 The abbreviations used are: LOH, loss of heterozygosity; RFLP, restriction fragment 2 To whom requests for reprints should be addressed, at Department of Pathology, length polymorphism; VNTR, variable number of tandem repeats; ATCC, American Type Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021. Culture Collection; PCR, polymerase chain reaction. 2848

parameters,@a,FNAaABLeASSâ€•D9S10'@&ageT@Table 1 AssociationbetweenLOH, microsatellite aiterations, andpathological

Ta @3 GO 1/2 0/0 1/3 1/1 1/2 1/2 1/3 T1 Wll 0/1 7/8 0/4 2111 0/3 2/6 3/12 3/lI 2W45Total11/613/1626/405/3913/655/2013/4113/6525/61o@1T2@W2 11/45Gil 3/141/1 17/29Q'l 5/341/2 9/491/2 3/140/1 lW320/2 9/491/2

2 1/9 Gfl 5/5 @5 3/10 1/4 1/6 5/Il 3/9 17/45Total11/603/1526f405/3812/634/1713/4113/6322/60N31/6 9f45WI 3/132/4 19/311/3 4/303/7 6/46WI 3/123/5 9/3046 4/46216

and MSt*usNo

3/10TotallG'532/1422/344/351N@,M'1W43 Wl01/10 1/417/27 5/72/26 2/99/45 2/145/15 tWl48/26 5/109/45 2/1318/43

1/595/2913/361 1/5821/53 @ a of allelic losses/number of informative cases. b Number of alterations/number of amplified cases.

as described previously (15). The dinuclcotide repeat polymorphism was presented LOH. There was no statistical significant correlation be anal@ by PCR amplification according to the Gene Amp kit protocol tween allelic losses at the IFNA and D9S3 loci and pathological (Perkin-Ehner Cetus, Norwalk, Cl) with the following modifications. The parameters. Only one case showed LOH for all the markers tested on final reaction volume was reduced to 20 p1; 2 g@Ci[a-32P]dCTPwere added 9p, indicating loss of the short arm without areas of retention. per reaction; and annealing temperatures ranged from 55 to 60Â°C,depending Chromosome 9q Allelic Losses. The 9q34.lâ€”34.3region was en the set of primers used. This was followed by polyaciylamide gel electro informed in terms of LOH at ASS, ABL, D9SJO, and/or D957 loci. phoresis on 8% acrylamide gels. For samples in which the normal tissue showed heterozygosity for a given probe or set of primers, the presence of the Seventy-one of 73 cases evaluated for this region were informative for alleles in the normal and the tumor tissues was assessed by densitometry using at least one of the probesused, obtainingan overall LOHrateof 45%. an Ultrascan XL Laser Densitoumter (Phannmia LKB Biotechnology, Pisca Allelic losses were distributed as follows: 2 of 2 Tis, 3 of 4 Ta, 6 of tawny, NJ). The relative decrease or the absence of the allele was established, 12 T1, and 21 of 53 T2@lesions. Data from each of the individual loci as reportedpreviously,comparingthenondeletedallelesin thenonnaland may be briefly summarized as follows. Thirteen of 65 informative tumor tissue counterpart (2, 3). cases showed LOH at the D9S7 locus. There was an association Statistical Methods. Fisher's exact test (16) was used to assess the asso between allelic losses at D9S7 and low tumor grade (P < 0.01), as ciation between clinicopathological factors and chromosome 9 alterations, including 9p and 9q allelic losses, as well as microsatellite abnormalities at chromosome 9 bcE Ch9 t.oci S Ntsratbons Results In the present study, 73 cases of primary bladder tumors were â€˜4 1 @ evaluated for LOH and microsatellite alterations on chromosome 9. D I (nâ€•65) 68% The probes and primerswere selected on the basis of chromosomal 11 J location and rate of polymorphism. Table 1 summarizes the results 13 obtained by RFLP and allelic losses identified by PCR allelotyping. A 12 -1 19% summary ofthe LOH and microsatellite alterations is shown in Fig. 1. :: J (n-48) Fig. 2 illustrates some of the altered patterns observed in this study.

LOH was defined by the complete or partial reduction in signal of one 12

of the polymorphicalleles. In the case of the microsatellitemarkers, 1$ alterations included allelic losses, expansions, and rearrangements.

Cases displaying microsatellite abnormalities were analyzed twice by 2L3 an additional PCR reaction and electrophoretic run. Overall, 56 of 73 211 13% (77%) cases showed chromosome 9 mutations. w (n-50) Chromosome 9p Allelic Losses. Sixty-five tumors with the cor $1 responding normal counterpart were analyzed by PCR allelotyping for D9S54 and JFNA loci. None of the superficial bladder tumors showed LOH for D9S54; however, 11 of 45 muscle invasive lesions showed 34.1 45% 342 (nâ€”73) allelic losses at this locus (P = 0.05). Twenty-six of 40 (65%) 342 informative cases presented LOH at the IFNA locus. These included 1 T11, 1 Ta, 7 T1, and 17 muscle invasive lesions (T2Ã·;Fig. 2). Fig. 1. DiagrammatiC represent*ion and summary of the frequency of chromosome 9 alterations in bladder tumors. The approximate physical location of each marker is Forty-eight cases were studied by RFLP for the D9S3 locus. Three indicated.@flupercentageofalterationrelatesto the numberof detectedmutationsper muscle invasive tumors of 16 (19%) informative cases analyzed informative cases studied. n, number of infonnmive cases studied with each marker. 2849

NT NT In order to better define the deleted region(s) on chromosome 9 in bladder cancer, we undertook the present study mapping the candidate NT regions using a set of highly polymorphic markers. The cases selected included both superficial and muscle invasive urothelial neoplasias. We observed that chromosome 9 alterations were indeed common events in bladder tumors. The highest frequency of allelic losses was found in the short arm of chromosome 9 with a 65% LOH in the IFNA gene cluster. This frequency increased to 80% when superficial le sions were considered as a subgroup. Detailed analysis at two other loci on this arm, D9554-telomeric and D953-centromeric to IFNA, indicated that only muscle invasive tumors had allelic losses in these regions. This suggests that a candidate tumor-suppressor gene on 9p D9S54 IFNa for certain superficial bladder tumors is closely linked to the JFNA locus. It has been reported that the locus for familial melanoma I susceptibility is in the chromosomal region 9pi3-p22 (18). In addi NT tion, genetic alterations near the interferon gene cluster have been documented in other human neoplasias, such as gliomas (19), non small cell lung cancer (20, 21), leukemias (22), and lymphomas (23).@ Allelic losses on the long arm of chromosome 9 were frequently observed, with an overall LOH of 45% for the 9q34.1â€”3 region. The frequency also increased to 56% when superficial tumors were ana D9S54 lyzed. Interestingly, only 13% of informative cases showed allelic losses at the D9S28 locus, centromerically located in reference to the D9S10 9q34.1â€”3region. Furthermore, it should be noted that all of these Fig. 2. Representative autoradiographies of alleic losses and alterations of chromo tumors were muscle invasive bladder cancers. These data indicate that some 9 as detected by Southern blots (D9SJO) or PCR amplification (D9S54 and IFNa). Note LOH (single arrow) and expansions (double arrows). N, normal; 7', tumor. a candidate tumor suppressor gene on 9q for a subset of superficial bladder tumors is located in the 9q34.1.-3 region. The galactosyl assessed by histopathological evaluation. D9S1O was informative in transferase that codes for the ABO histoblood group-related antigens 41 of 65 (63%) cases (Fig. 2). Thirteen of these informative samples maps to this area (24). Altered patterns of expression of these anti revealed LOH, but there was no correlation with pathological param genic determinants has been reported as a common phenotype in eters. Sixty-eight cases were analyzed for the ABL locus by RFLP and bladder cancer (25, 26). More recently, it has been reported that PCR allelotyping. Thirteen of 65 informative cases showed allelic the target gene for the Gorlin syndrome maps to chromosome losses. For the ASS locus, 14 cases were analyzed by PCR allelotyping 9q22.3â€”q31(27, 28). This is an autosomal dominant disorder predis and 56 by RFLP. Five of 20 informative cases showed LOH at this posing to basal cell carcinomas of the skin and other neoplasms, locus. Finally, 50 samples were tested on the D9528 locus. Only 5 including bladder tumors. muscle invasive tumors of 39 (13%) informative cases presented LOH Allelic losses reported here on 9p and 9q are in concordance with at this site, whereas none of the 5 informative superficial tumors previous findings from other groups (5, 6, 8â€”10).In addition, we showed allelic losses on this locus. observed that 68% of 9p LOH samples showed losses for 9q. Microsatellite Alterations. The D9554 locus was amplified by However, Ta lesions appeared to have 9q LOH as the main genetic PCR in 61 tumors. Twenty-five (41%) cases showed alterations, alteration, whereas T1 tumors presented with a mixture of aberrant including 13 expansions and 1 rearrangement, in addition to the 11 chromosome 9 genotypes (i.e., 9p-, 9q-, and 9p-/9q-). Moreover, allelic losses described above. The expansions occurred in one Ta, one all allelic losses observed at D9554 occurred in muscle invasive T15,three T1, and eight T2@lesions. A muscle invasive case showed tumors. Taken together, these data suggest that 9p LOH may be a rearrangement at the D9554 locus. No microsatellite alterations at associated with a more aggressive biological behavior or early the ASS locus were observed of 14 amplified cases. We found only 2 disease progression. of 48 amplified samples with expansions at the ABL locus. They both We also designed this study to evaluate the existence and frequency showed concomitant alterations at D9S54, corresponding to an expan of microsatellite alterations occurring on chromosome 9 in urothelial sion and an allelic loss at this site. neoplasias. These abnormalities have been associated with the repli cation error phenotype, a reflection of genetic instability (12â€”14).The Discussion ASS microsatellite displayed normal patterns in all amplified samples, Loss of genetic material on chromosome 9 is an early abnormality whereas two expansions were identified at the ABL locus. However, detected in bladder tumors. Monosomy of this chromosome and allelic we found that 41% of the amplified cases for the D9S54 locus losses of 9q have been reported as frequent alterations in low-grade, exhibited instability, indicating that this is not a random event in low-stage bladder cancer (1, 3â€”7).Due to these observations, it has bladder cancer. The alterations characterized on D9S54 did not seem been suggested that this event associates with the development of to relate to 9p and/or 9q status. Gonzalez-Zulueta et a!. (1 1) have papillary superficial bladder tumors (3, 5, 7). Furthermore, the diver recently documented somatic instability at microsatellite repeats on gence in the percentage of chromosome 9 allelic losses observed chromosome 9 in four bladder tumors (11). between papillary superficial tumors and muscle invasive lesions has In the present study and considering all types of genetic alterations, been the basis for the working hypothesis that two genetic pathways we found that 77% of the informative cases had chromosome 9 characterize the evolution of bladder cancer (3, 17). More recently, the mutations, representing the largest percentage documented to date. existence of two altered loci, one in each of both chromosomal arms,

has been postulated (10). 4 R. Chaganti et aL, personal communication. 2850

This may be due to the combination of assays used as well as the loci 12. Leach, F. S., Nicolaides, N. C., Papadopoulus, N., Liu, B., and Vogelstein, B. and regions analyzed. It is possible that the cohort of patients selected, Mutations of a mutS homolog in hereditary nonpolyposis colorectal cancer. Cell, 75: 1215â€”1225,1993. based on histopathology evaluation and good tissue preservation, may 13. Peltomaki, P., Lothe, R. A., Aaltonen, L. A., Pylkkanen, L., Nystrom-Lahti, M., have also contributed to the results reported here. Further studies Seruca, R., David, L, HoIm, R., Ryberg, D., Haugen, A., Brogger, A., Borresen, centering on the role of these abnormalities, mainly as they occur in A. L.,andde IaChapelle,A.Microsatelliteinstabilityisassociatedwithtumorsthat characterize the hereditary nonpolyposis colorectal carcinoma syndrome. Cancer a large and well-defined group of superficial bladder tumors, will be Rex., 53: 5853â€”5855,1993. needed in order to better understand their involvement as either 14. Ionov, Y., Peinado, M. A., Malkhosyan, S., Shibata. D., and Perucho, M. Ubiquitous primary or secondary events in urothelial neoplasias. somatic mutations in simple repeated sequences reveal a new mechanism for colonic carcinogenesis. Nature (Lond.), 363: 558â€”561, 1993. 15. Dalbagni, G., Presti, J. C. J., Reuter, V. E., Thang, Z. F., Sarkis, A. S., Fair, W. R., Acknowledgments and Cordon-Cardo, C. Molecular genetic alterations of chromosome 17 and p53 nuclear overexpression in human bladder cancer. Diagn. Mol. Pathol., 2: 4â€”13,1993. We thank Des. R. Chaganti, M. Diaz, M. Perucho, J. Rosai, and Zuo-Feng 16. Metha, C. R., and Patel, N. R. A network algorithm for performing Fisher's exact test Zhang for their helpful suggestions regarding this study and their critical in r X c contingency tables. J. Am. Statist. Assoc., 78: 427â€”434, 1983. review of the manuscript. 17. Spruck, C. H. I., et a!. Two molecular pathways to transitional cell carcinoma of the bladder. Cancer Res., 54: 784â€”788,1994. 18. Cannon-Albright, L. A., el aL Assignment of a locus for familial melanoma, MLM, References to chromosome 9pl3-p22. Science (Washington DC), 258: 1148â€”1152,1992. 1. Gibas, Z., Prout, G. R., Connolly, J. 0., Pontis, J. E., and Sandberg, A. A. Nonrandom 19. James, C. D., He, J., Carlbom, E., Nordenskjold, M., Cavenee, W. K., and Collins, chmmosomal changes in transitional cell carcinoma of the bladder. Cancer Res., 44: V. P. Chromosome9deletionmappingrevealsinterferona andinterferon @-lgene 1257â€”1264,1984. deletions in human glial tumors. Cancer Res., 51: 1684â€”1688, 1991. 2. Presti, J. C. J., Reuter, V. E., Galan, T., Fair, W. R., and Cordon-Cardo, C. Molecular 20. Olopade, 0. I., Buchhagen, D. L, Malik, K., Sherman, J., Nobori, T., Bader, S., Nau, genetic alterations in superficial and locally advanced human bladder cancer. Cancer M. M.,Gazda,A. F., Minna,J. D., and Diaz,M. 0. Homozygousdeletionof the Res., 51: 5405â€”5409,1991. interferon genes defines the critical region on 9p that is deleted in lung cancers. 3. Dalbagni, ci., Presti, J., Reuter, V., Fair, W. R., and Cordon-Cardo, C. Genetic Cancer Res., 53: 2410â€”2415, 1993. alterations in bladder cancer. Lancet, 342: 469â€”471, 1993. 21. Merlo, A., Gabrielson, E., Askin, F., and Sidransky, D. Frequent loss of chromosome 4. Knowles, M. A., Elder, P. A., Williamson, M., Cairns, J. P., Shaw, M. E., and Law, 9 in human primary non-small cell lung cancer. Cancer Res., 54: 640â€”642, 1994. M.G. Allelotypeofhumanbladdercancer.CancerRes.,54: 531â€”538,1994. 22. Diaz, M. 0., Ziemin, S., La Beau, M. M., Pitha, P. M., Smith, S. D., Chilcote, R. R., 5. Miyao, N., Tsai, Y. C., Lerner, S. P., Olumi, A. F., Spruck, C. H. I., Gonzalez and Rowley, J. D. Homozygous deletion of the a- and @3-1interferongenes in human Zulueta, M., Nichols, P. W., Skinner, D. G., and Jones, P. A. Role of chromosome leukemia and derived cell lines. Proc. NatI. Acad. Sci. USA, 85: 5259â€”5263, 1988. 9 in humanbladdercancer.CancerRes.,53: 4066â€”4070,1993. 23. Olopade, 0. 1.,Bohlander, S. K., Pomykala, H., Maltepe, E., Van Melle, E., La beau, 6. Cairns, P., Shaw, M. E., and Knowles, M. A. Preliminary mapping of the deleted M.M.,andDiaz,M.0. Mappingoftheshortestregionofoverlapofdeletionsofthe region of chromosome 9 in bladder cancer. Cancer Res., 53: 1230â€”1232, 1993. short arm of chromosome 9 associated with human neoplasia. Genomics, 14: 7. Caims, P., Shaw, M. E., and Knowles, M. A. Initiation of bladder cancer may involve 437â€”443,1992. deletionof a tumor-suppressorgeneon chromosome9.Oncogene,8: 1083-1085, 24. Yamamoto, F. I., Clausen, H., White, T., Marker, J., and Hakomori, S. Molecular 1993. genetic basis of the histo-blood group ABO system. Nature (Lond.). 345: 229â€”233, 8. Tsai, Y. C., Nichols, P. W., Hiti, A. L., Williams, Z., Skinner, D., and Jones, P. A. 1990. Allelic losses of chromosomes 9, 11, and 17 in human bladder cancer. Cancer Res., 25. Cordon-Cardo, C., Reuter, V. E., Uoyd, K. 0., Sheinfel, i., Fair, W. R., Old, L. J., 50: 44â€”47,1990. Memlamed, M. R. Blood group-related antigens in human urothelium: enhanced 9. Habuchi, T., Ogawa, 0., Kakehi, Y., Ogura, K., Koshiba, M., Hamazaki, S., Taka expression of precursor, LaX and LaY determinants in urothelial carcinoma. Cancer hashi, R., Sugiyama, T., and Yoshida, 0. Accumulated allelic losses in the develop Res.,48:4113â€”4120,1988. ment of invasive urothelial cancer.. Int. J. Cancer, 53: 579â€”584,1993. 26. Orntoft, T. F., Wolf, H., and Watkins, W. M. Activity of the human blood group 10. Ruppert,J. M., Tokino,K., and Sidransky,D. Evidencefor two bladdercancer ABO, Se, H, 12, and X gene-encoded glycosyltransferases in normal and malignant supressor loci on human chromosome 9. Cancer Res., 53: 5093â€”5095,1993. bladder urothelium. Cancer Res., 48: 4427â€”4433,1988. 11. Gonzalez-Zulueta,M.,Ruppert,J. M., Tokino,K., Tsai, Y. C., Spruck,C. H. I., 27. Farndon, P. A., Del Mastro, R. 0., Evans, D. G. R., and Kilpatrick, M. W. Location Miyao, N., Nichols, P. W., Hermann, 0. 0., Horn, T., Steven, K., Summerhayes, of gene for Gorlin syndrome. Lancet, 339: 581â€”582,1992. I. C., Sidransky,D.,and Jones,P. A. Microsatelliteinstabilityin bladdercancer. 28. Gailani,M.R.,et aLDevelopmentaldefectsinGorlinsyndromerelatedtoa putative CancerRes.,53: 5620â€”5623,1993. tumor suppressor gene on chromosome 9. Cell, 69: 111â€”117,1992.

2851

Irene Orlow, Pilar Lianes, Louis Lacombe, et al.

Cancer Res 1994;54:2848-2851.

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