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. 2001 Feb;84(3):321–328. doi: 10.1054/bjoc.2000.1595

Immunohistochemical analysis of expression and allelotype of mismatch repair genes (hMLH1 and hMSH2) in bladder cancer

H Sh Kassem 1,3, J M Varley 2, S M Hamam 3, G P Margison 1
PMCID: PMC2363732  PMID: 11161395

Abstract

Mutation of human homologues of DNA mismatch repair (MMR) genes in tumours has been shown to be associated with the phenomenon of microsatellite instability (MSI). Several studies have reported the occurrence of MSI in bladder cancer, but evidence of involvement of MMR genes in the pathogenesis of this cancer is still unclear. We therefore utilized quantitative immunohistochemical (IHC) image analysis and PCR-based allelotype analysis to determine hMLH1 and hMSH2 genes alteration in a cohort of Egyptian bladder cancer samples. IHC analysis of 24 TCC and 12 SCC revealed marked- intra and intertumour heterogeneity in the levels of expression of the two MMR proteins. One TCC lost MLH1 expression and one lost MSH2, (1/24, 4%), and one SCC lost MSH2 (1/12, 8%). A large proportion of analysed tumours revealed a percentage positivity of less than 50% for MLH1 and MSH2 expression (44% and 69%, respectively). Complete loss of heterozygosity in three dinucleotide repeats lying within, or in close proximity to, hMLH1 and hMSH2 was rare (2/57, (4%) for MLH1; and 1/55, (2%) for MSH2), however allelic imbalance was detected in 11/57 (hMLH1) and 10/55 (hMSH2) at any of the informative microsatellite loci. These alterations in structure and expression of DNA MMR genes suggest their possible involvement in the tumorigenesis and/or progression of bladder cancer. © 2001 Cancer Research Campaign http://www.bjcancer.com

Keywords: MLH1, MSH2, allelotype, immunohistochemistry, bladder cancer

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Selected References

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  1. Aaltonen L. A., Peltomäki P., Leach F. S., Sistonen P., Pylkkänen L., Mecklin J. P., Järvinen H., Powell S. M., Jen J., Hamilton S. R. Clues to the pathogenesis of familial colorectal cancer. Science. 1993 May 7;260(5109):812–816. doi: 10.1126/science.8484121. [DOI] [PubMed] [Google Scholar]
  2. Aaltonen L. A., Peltomäki P., Mecklin J. P., Järvinen H., Jass J. R., Green J. S., Lynch H. T., Watson P., Tallqvist G., Juhola M. Replication errors in benign and malignant tumors from hereditary nonpolyposis colorectal cancer patients. Cancer Res. 1994 Apr 1;54(7):1645–1648. [PubMed] [Google Scholar]
  3. Aarnio M., Sankila R., Pukkala E., Salovaara R., Aaltonen L. A., de la Chapelle A., Peltomäki P., Mecklin J. P., Järvinen H. J. Cancer risk in mutation carriers of DNA-mismatch-repair genes. Int J Cancer. 1999 Apr 12;81(2):214–218. doi: 10.1002/(sici)1097-0215(19990412)81:2<214::aid-ijc8>3.0.co;2-l. [DOI] [PubMed] [Google Scholar]
  4. Benachenhou N., Guiral S., Gorska-Flipot I., Labuda D., Sinnett D. Frequent loss of heterozygosity at the DNA mismatch-repair loci hMLH1 and hMSH3 in sporadic breast cancer. Br J Cancer. 1999 Mar;79(7-8):1012–1017. doi: 10.1038/sj.bjc.6690162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Benachenhou N., Guiral S., Gorska-Flipot I., Labuda D., Sinnett D. High resolution deletion mapping reveals frequent allelic losses at the DNA mismatch repair loci hMLH1 and hMSH3 in non-small cell lung cancer. Int J Cancer. 1998 Jul 17;77(2):173–180. doi: 10.1002/(sici)1097-0215(19980717)77:2<173::aid-ijc1>3.0.co;2-n. [DOI] [PubMed] [Google Scholar]
  6. Benachenhou N., Guiral S., Gorska-Flipot I., Michalski R., Labuda D., Sinnett D. Allelic losses and DNA methylation at DNA mismatch repair loci in sporadic colorectal cancer. Carcinogenesis. 1998 Nov;19(11):1925–1929. doi: 10.1093/carcin/19.11.1925. [DOI] [PubMed] [Google Scholar]
  7. Boring C. C., Squires T. S., Tong T., Montgomery S. Cancer statistics, 1994. CA Cancer J Clin. 1994 Jan-Feb;44(1):7–26. doi: 10.3322/canjclin.44.1.7. [DOI] [PubMed] [Google Scholar]
  8. Brentnall T. A. Microsatellite instability. Shifting concepts in tumorigenesis. Am J Pathol. 1995 Sep;147(3):561–563. [PMC free article] [PubMed] [Google Scholar]
  9. Brodsky G. L. Pathology of bladder carcinoma. Hematol Oncol Clin North Am. 1992 Feb;6(1):59–80. [PubMed] [Google Scholar]
  10. Bronner C. E., Baker S. M., Morrison P. T., Warren G., Smith L. G., Lescoe M. K., Kane M., Earabino C., Lipford J., Lindblom A. Mutation in the DNA mismatch repair gene homologue hMLH1 is associated with hereditary non-polyposis colon cancer. Nature. 1994 Mar 17;368(6468):258–261. doi: 10.1038/368258a0. [DOI] [PubMed] [Google Scholar]
  11. Chong J. M., Fukayama M., Hayashi Y., Takizawa T., Koike M., Konishi M., Kikuchi-Yanoshita R., Miyaki M. Microsatellite instability in the progression of gastric carcinoma. Cancer Res. 1994 Sep 1;54(17):4595–4597. [PubMed] [Google Scholar]
  12. Christensen M., Jensen M. A., Wolf H., Orntoft T. F. Pronounced microsatellite instability in transitional cell carcinomas from young patients with bladder cancer. Int J Cancer. 1998 Aug 21;79(4):396–401. doi: 10.1002/(sici)1097-0215(19980821)79:4<396::aid-ijc15>3.0.co;2-3. [DOI] [PubMed] [Google Scholar]
  13. Cox E. C. Bacterial mutator genes and the control of spontaneous mutation. Annu Rev Genet. 1976;10:135–156. doi: 10.1146/annurev.ge.10.120176.001031. [DOI] [PubMed] [Google Scholar]
  14. Cox E. C. Mutator gene studies in Escherichia coli: the mutT gene. Genetics. 1973 Apr;73(Suppl):67–80. [PubMed] [Google Scholar]
  15. Fink D., Nebel S., Aebi S., Zheng H., Kim H. K., Christen R. D., Howell S. B. Expression of the DNA mismatch repair proteins hMLH1 and hPMS2 in normal human tissues. Br J Cancer. 1997;76(7):890–893. doi: 10.1038/bjc.1997.480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fishel R., Lescoe M. K., Rao M. R., Copeland N. G., Jenkins N. A., Garber J., Kane M., Kolodner R. The human mutator gene homolog MSH2 and its association with hereditary nonpolyposis colon cancer. Cell. 1994 Apr 8;77(1):1–166. [PubMed] [Google Scholar]
  17. Gonzalez-Zulueta M., Ruppert J. M., Tokino K., Tsai Y. C., Spruck C. H., 3rd, Miyao N., Nichols P. W., Hermann G. G., Horn T., Steven K. Microsatellite instability in bladder cancer. Cancer Res. 1993 Dec 1;53(23):5620–5623. [PubMed] [Google Scholar]
  18. Gyapay G., Morissette J., Vignal A., Dib C., Fizames C., Millasseau P., Marc S., Bernardi G., Lathrop M., Weissenbach J. The 1993-94 Généthon human genetic linkage map. Nat Genet. 1994 Jun;7(2 Spec No):246–339. doi: 10.1038/ng0694supp-246. [DOI] [PubMed] [Google Scholar]
  19. Habano W., Sugai T., Nakamura S. Mismatch repair deficiency leads to a unique mode of colorectal tumorigenesis characterized by intratumoral heterogeneity. Oncogene. 1998 Mar 12;16(10):1259–1265. doi: 10.1038/sj.onc.1201651. [DOI] [PubMed] [Google Scholar]
  20. Han H. J., Yanagisawa A., Kato Y., Park J. G., Nakamura Y. Genetic instability in pancreatic cancer and poorly differentiated type of gastric cancer. Cancer Res. 1993 Nov 1;53(21):5087–5089. [PubMed] [Google Scholar]
  21. Hemminki A., Peltomäki P., Mecklin J. P., Järvinen H., Salovaara R., Nyström-Lahti M., de la Chapelle A., Aaltonen L. A. Loss of the wild type MLH1 gene is a feature of hereditary nonpolyposis colorectal cancer. Nat Genet. 1994 Dec;8(4):405–410. doi: 10.1038/ng1294-405. [DOI] [PubMed] [Google Scholar]
  22. Hsu S. M., 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 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  23. Jin T. X., Furihata M., Yamasaki I., Kamada M., Liang S. B., Ohtsuki Y., Shuin T. Human mismatch repair gene (hMSH2) product expression in relation to recurrence of transitional cell carcinoma of the urinary bladder. Cancer. 1999 Jan 15;85(2):478–484. [PubMed] [Google Scholar]
  24. Kane M. F., Loda M., Gaida G. M., Lipman J., Mishra R., Goldman H., Jessup J. M., Kolodner R. Methylation of the hMLH1 promoter correlates with lack of expression of hMLH1 in sporadic colon tumors and mismatch repair-defective human tumor cell lines. Cancer Res. 1997 Mar 1;57(5):808–811. [PubMed] [Google Scholar]
  25. Kolodner R. D. Mismatch repair: mechanisms and relationship to cancer susceptibility. Trends Biochem Sci. 1995 Oct;20(10):397–401. doi: 10.1016/s0968-0004(00)89087-8. [DOI] [PubMed] [Google Scholar]
  26. Lairmore T. C., Norton J. A. Advances in molecular genetics. Am J Surg. 1997 Jan;173(1):37–43. doi: 10.1016/S0002-9610(96)00363-7. [DOI] [PubMed] [Google Scholar]
  27. Leach F. S., Nicolaides N. C., Papadopoulos N., Liu B., Jen J., Parsons R., Peltomäki P., Sistonen P., Aaltonen L. A., Nyström-Lahti M. Mutations of a mutS homolog in hereditary nonpolyposis colorectal cancer. Cell. 1993 Dec 17;75(6):1215–1225. doi: 10.1016/0092-8674(93)90330-s. [DOI] [PubMed] [Google Scholar]
  28. Leach F. S., Polyak K., Burrell M., Johnson K. A., Hill D., Dunlop M. G., Wyllie A. H., Peltomaki P., de la Chapelle A., Hamilton S. R. Expression of the human mismatch repair gene hMSH2 in normal and neoplastic tissues. Cancer Res. 1996 Jan 15;56(2):235–240. [PubMed] [Google Scholar]
  29. Leung S. Y., Yuen S. T., Chung L. P., Chu K. M., Chan A. S., Ho J. C. hMLH1 promoter methylation and lack of hMLH1 expression in sporadic gastric carcinomas with high-frequency microsatellite instability. Cancer Res. 1999 Jan 1;59(1):159–164. [PubMed] [Google Scholar]
  30. Li M., Zhang Z. F., Reuter V. E., Cordon-Cardo C. Chromosome 3 allelic losses and microsatellite alterations in transitional cell carcinoma of the urinary bladder. Am J Pathol. 1996 Jul;149(1):229–235. [PMC free article] [PubMed] [Google Scholar]
  31. Lim P. C., Tester D., Cliby W., Ziesmer S. C., Roche P. C., Hartmann L., Thibodeau S. N., Podratz K. C., Jenkins R. B. Absence of mutations in DNA mismatch repair genes in sporadic endometrial tumors with microsatellite instability. Clin Cancer Res. 1996 Nov;2(11):1907–1911. [PubMed] [Google Scholar]
  32. Liu B., Parsons R. E., Hamilton S. R., Petersen G. M., Lynch H. T., Watson P., Markowitz S., Willson J. K., Green J., de la Chapelle A. hMSH2 mutations in hereditary nonpolyposis colorectal cancer kindreds. Cancer Res. 1994 Sep 1;54(17):4590–4594. [PubMed] [Google Scholar]
  33. Lo Muzio L., Nocini P., Mignogna M. D., Pannone G., Staibano S., Procaccini M., Mariggio M. A., Dolci M., De Rosa G. Immunocytochemical detection of hMSH2 and hMLH1 expression in oral SCC. Anticancer Res. 1999 Mar-Apr;19(2A):933–940. [PubMed] [Google Scholar]
  34. Loeb L. A. Microsatellite instability: marker of a mutator phenotype in cancer. Cancer Res. 1994 Oct 1;54(19):5059–5063. [PubMed] [Google Scholar]
  35. Loeb L. A. Mutator phenotype may be required for multistage carcinogenesis. Cancer Res. 1991 Jun 15;51(12):3075–3079. [PubMed] [Google Scholar]
  36. Mao L. Genetic alterations as clonal markers for bladder cancer detection in urine. J Cell Biochem Suppl. 1996;25:191–196. [PubMed] [Google Scholar]
  37. Mao L., Lee D. J., Tockman M. S., Erozan Y. S., Askin F., Sidransky D. Microsatellite alterations as clonal markers for the detection of human cancer. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9871–9875. doi: 10.1073/pnas.91.21.9871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Mao L., Schoenberg M. P., Scicchitano M., Erozan Y. S., Merlo A., Schwab D., Sidransky D. Molecular detection of primary bladder cancer by microsatellite analysis. Science. 1996 Feb 2;271(5249):659–662. doi: 10.1126/science.271.5249.659. [DOI] [PubMed] [Google Scholar]
  39. Modrich P. Mechanisms and biological effects of mismatch repair. Annu Rev Genet. 1991;25:229–253. doi: 10.1146/annurev.ge.25.120191.001305. [DOI] [PubMed] [Google Scholar]
  40. Mourah S., Cussenot O., Vimont V., Desgrandchamps F., Teillac P., Cochant-Priollet B., Le Duc A., Fiet J., Soliman H. Assessment of microsatellite instability in urine in the detection of transitional-cell carcinoma of the bladder. Int J Cancer. 1998 Dec 18;79(6):629–633. doi: 10.1002/(sici)1097-0215(19981218)79:6<629::aid-ijc13>3.0.co;2-1. [DOI] [PubMed] [Google Scholar]
  41. Nicolaides N. C., Papadopoulos N., Liu B., Wei Y. F., Carter K. C., Ruben S. M., Rosen C. A., Haseltine W. A., Fleischmann R. D., Fraser C. M. Mutations of two PMS homologues in hereditary nonpolyposis colon cancer. Nature. 1994 Sep 1;371(6492):75–80. doi: 10.1038/371075a0. [DOI] [PubMed] [Google Scholar]
  42. Orlow I., Lianes P., Lacombe L., Dalbagni G., Reuter V. E., Cordon-Cardo C. Chromosome 9 allelic losses and microsatellite alterations in human bladder tumors. Cancer Res. 1994 Jun 1;54(11):2848–2851. [PubMed] [Google Scholar]
  43. Papadopoulos N., Nicolaides N. C., Wei Y. F., Ruben S. M., Carter K. C., Rosen C. A., Haseltine W. A., Fleischmann R. D., Fraser C. M., Adams M. D. Mutation of a mutL homolog in hereditary colon cancer. Science. 1994 Mar 18;263(5153):1625–1629. doi: 10.1126/science.8128251. [DOI] [PubMed] [Google Scholar]
  44. Parkin D. M., Pisani P., Ferlay J. Estimates of the worldwide incidence of eighteen major cancers in 1985. Int J Cancer. 1993 Jun 19;54(4):594–606. doi: 10.1002/ijc.2910540413. [DOI] [PubMed] [Google Scholar]
  45. Parsons R., Li G. M., Longley M. J., Fang W. H., Papadopoulos N., Jen J., de la Chapelle A., Kinzler K. W., Vogelstein B., Modrich P. Hypermutability and mismatch repair deficiency in RER+ tumor cells. Cell. 1993 Dec 17;75(6):1227–1236. doi: 10.1016/0092-8674(93)90331-j. [DOI] [PubMed] [Google Scholar]
  46. Parsons R., Li G. M., Longley M., Modrich P., Liu B., Berk T., Hamilton S. R., Kinzler K. W., Vogelstein B. Mismatch repair deficiency in phenotypically normal human cells. Science. 1995 May 5;268(5211):738–740. doi: 10.1126/science.7632227. [DOI] [PubMed] [Google Scholar]
  47. Peltomäki P. DNA mismatch repair gene mutations in human cancer. Environ Health Perspect. 1997 Jun;105 (Suppl 4):775–780. doi: 10.1289/ehp.105-1470030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Peltomäki P., de la Chapelle A. Mutations predisposing to hereditary nonpolyposis colorectal cancer. Adv Cancer Res. 1997;71:93–119. doi: 10.1016/s0065-230x(08)60097-4. [DOI] [PubMed] [Google Scholar]
  49. Risinger J. I., Berchuck A., Kohler M. F., Watson P., Lynch H. T., Boyd J. Genetic instability of microsatellites in endometrial carcinoma. Cancer Res. 1993 Nov 1;53(21):5100–5103. [PubMed] [Google Scholar]
  50. Rosin M. P., Cairns P., Epstein J. I., Schoenberg M. P., Sidransky D. Partial allelotype of carcinoma in situ of the human bladder. Cancer Res. 1995 Nov 15;55(22):5213–5216. [PubMed] [Google Scholar]
  51. Steiner G., Schoenberg M. P., Linn J. F., Mao L., Sidransky D. Detection of bladder cancer recurrence by microsatellite analysis of urine. Nat Med. 1997 Jun;3(6):621–624. doi: 10.1038/nm0697-621. [DOI] [PubMed] [Google Scholar]
  52. Thibodeau S. N., Bren G., Schaid D. Microsatellite instability in cancer of the proximal colon. Science. 1993 May 7;260(5109):816–819. doi: 10.1126/science.8484122. [DOI] [PubMed] [Google Scholar]
  53. Thibodeau S. N., French A. J., Roche P. C., Cunningham J. M., Tester D. J., Lindor N. M., Moslein G., Baker S. M., Liskay R. M., Burgart L. J. Altered expression of hMSH2 and hMLH1 in tumors with microsatellite instability and genetic alterations in mismatch repair genes. Cancer Res. 1996 Nov 1;56(21):4836–4840. [PubMed] [Google Scholar]
  54. Tomlinson I. P., Ilyas M., Bodmer W. F. Allele loss occurs frequently at hMLH1, but rarely at hMSH2, in sporadic colorectal cancers with microsatellite instability. Br J Cancer. 1996 Nov;74(10):1514–1517. doi: 10.1038/bjc.1996.582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Uchida T., Wang C., Wada C., Iwamura M., Egawa S., Koshiba K. Microsatellite instability in transitional cell carcinoma of the urinary tract and its relationship to clinicopathological variables and smoking. Int J Cancer. 1996 Apr 22;69(2):142–145. doi: 10.1002/(SICI)1097-0215(19960422)69:2<142::AID-IJC13>3.0.CO;2-B. [DOI] [PubMed] [Google Scholar]
  56. Varley J. M., McGown G., Thorncroft M., James L. A., Margison G. P., Forster G., Evans D. G., Harris M., Kelsey A. M., Birch J. M. Are there low-penetrance TP53 Alleles? evidence from childhood adrenocortical tumors. Am J Hum Genet. 1999 Oct;65(4):995–1006. doi: 10.1086/302575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Wada C., Shionoya S., Fujino Y., Tokuhiro H., Akahoshi T., Uchida T., Ohtani H. Genomic instability of microsatellite repeats and its association with the evolution of chronic myelogenous leukemia. Blood. 1994 Jun 15;83(12):3449–3456. [PubMed] [Google Scholar]
  58. Wieland I., Ammermüller T., Böhm M., Totzeck B., Rajewsky M. F. Microsatellite instability and loss of heterozygosity at the hMLH1 locus on chromosome 3p21 occur in a subset of nonsmall cell lung carcinomas. Oncol Res. 1996;8(1):1–5. [PubMed] [Google Scholar]

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