Skip to main content
PMC home page
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 2002 Jul;93(7):767–773. doi: 10.1111/j.1349-7006.2002.tb01318.x

DNA Hypermethylation Status of Multiple Genes in Prostate Adenocarcinomas

Noboru Konishi 1,✉, Mitsutoshi Nakamura 1, Munehiro Kishi 1, Masayoshi Nishimine 1, Eiwa Ishida 1, Keiji Shimada 1
PMCID: PMC5927069  PMID: 12149142

Abstract

Multiple genetic mutations and epigenetic methylation are believed to be involved in prostate carcinogenesis, but it is not known whether these events are independent or correlated in some fashion. We therefore studied 32 prostate adenocarcinomas not only for deletions and/or mutations of multiple suspect genes, but also for aberrant DNA methylation using methylation‐specific PCR (MSP). Of those genes examined, p16INK4a, O6‐MGMT, and GST‐P were found to be the most frequently methylated (66%, 25% and 75% of cases, respectively), while methylations of p14ARF, RB1, p21waf1, and p27Kip1 were far less common (3%, 6%, 6% and 6% of cases, respectively). Methylation of O6‐MGMT and GST‐P genes was defective in about 19% of the cases and there were occasional simultaneous deletions and methylations of p14ARF and p16INK4a genes (13% and 3% of cases, respectively). In p16INK4a, methylation occurred in the promoter region in 9% of samples and in exon 2 in 66% of tumors. Hypermethylation of O6‐MGMT with concurrent p53 and ras gene mutations were found in 6% and 13% of specimens, respectively; among those tumors with high Gleason scores were 2 carcinomas showing hypermethylated O6‐MGMT with G‐to‐A transitions in K‐ras. Our results demonstrate that multiple genes of a subset common in prostate carcinomas are methylated and not infrequently show concurrent deletions. Further, there is a suggestion that specific combinations of hypermethylation and mutation correlate to tumor malignancy.

Keywords: Prostate carcinoma, Methylation, Mutation, Gene

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

REFERENCES

  • 1. ) Rennie , P. S. and Nelson , C. C.Epigenetic mechanisms for progression of prostate cancer . Cancer Metastasis Rev. , 17 , 401 – 409 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) Herman , J. G. , Graff , J. R. , Myohanen , S. , Nelkin , B. D. and Baylin , S. B.Methylation‐specific PCR: a novel PCR assay for methylation status of CpG islands . Proc. Natl. Acad. Sci. USA , 93 , 9821 – 9826 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3. ) Millar , D. S. , Ow , K. K. , Paul , C. L. , Russell , P. J. , Molloy , P. L. and Clark , S. J.Detailed methylation analysis of the glutathione S‐transferase π (GSTP1) gene in prostate cancer . Oncogene , 18 , 1313 – 1324 ( 1999. ). [DOI] [PubMed] [Google Scholar]
  • 4. ) Xing , E. P. , Nie , Y. , Wang , L. D. , Yang , G. Y. and Yang , C. S.Aberrant methylation of p16INK4a and deletion of pl51NK4b are frequent events in human esophageal cancer in Linxian, China . Carcinogenesis , 20 , 77 – 84 ( 1999. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Cairns , P. , Tokino , K. , Eby , Y. and Sidransky , D.Homozygous deletions of 9p21 in primary human bladder tumors detected by comparative multiplex polymerase chain reaction . Cancer Res. , 54 , 1422 – 1424 ( 1994. ). [PubMed] [Google Scholar]
  • 6. ) Gleason , D. F.Histologic grading of prostate cancer: a perspective . Hum. Pathol , 23 , 273 – 279 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Konishi , N. , Hiasa , Y. , Matsuda , H. , Tao , M. , Tsuzuki , T. , Hayashi , I. , Kitahori , Y. , Shiraishi , T. , Yatani , R. , Shimazaki , J. and Lin , J.‐C.Intratumor cellular heterogeneity and alterations in ras oncogene and p53 tumor suppressor gene in human prostate carcinoma . Am. J. Pathol. , 147 , 1112 – 1122 ( 1995. ). [PMC free article] [PubMed] [Google Scholar]
  • 8. ) Nakamura , M. , Yonekawa , Y. , Kleihues , P. and Ohgaki , H.Promoter hypermethylation of the RB1 gene in glioblastomas . Lab. Invest. , 81 , 77 – 82 ( 2001. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Nakamura , M. , Watanabe , T. , Klangby , U. , Asker , C. , Wiman , K. , Yonekawa , Y. , Kleihues , P. and Ohgaki , H.p14ARF deletion and methylation in genetic pathways to glioblastomas . Brain Pathol. , 11 , 159 – 168 ( 2001. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10. ) Corn , P. G. , Kuerbitz , S. J. , van Noesel , M. M. , Esteller , M. , Compitello , N. , Baylin , S. B. and Herman , J. G.Transcriptional silencing of the p73 gene in acute lymphoblastic leukemia and Burkitt's lymphoma is associated with 5′ CpG island methylation . Cancer Res. , 59 , 3352 – 3356 ( 1999. ). [PubMed] [Google Scholar]
  • 11. ) Esteller , M. , Hamilton , S. R. , Burger , P. C. , Baylin , S. B. and Herman , J. G.Inactivation of the DNA repair gene O6‐methylguanine‐DNA methyltransferase by promoter hypermethylation is a common event in primary human neoplasia . Cancer Res. , 59 , 793 – 797 ( 1999. ). [PubMed] [Google Scholar]
  • 12. ) Esteller , M. , Sanchez‐Cespedes , M. , Rosell , R. , Sidransky , D. , Baylin , S. B. and Herman , J. G.Detection of aberrant promoter hypermethylation of tumor suppressor genes in serum DNA from non‐small cell lung cancer patients . Cancer Res. , 59 , 67 – 70 ( 1999. ). [PubMed] [Google Scholar]
  • 13. ) Esteller , M. , Tortola , S. , Toyota , M. , Capella , G. , Peinado , M. A. , Baylin , S. B. and Herman , J. G.Hypermethylation‐associated inactivation of p14ARF is independent of p16INK4amethylation and p53 mutational status . Cancer Res. , 60 , 129 – 133 ( 2000. ). [PubMed] [Google Scholar]
  • 14. ) Esteller , M. , Toyota , M. , Sanchez‐Cespedes , M. , Capella , G. , Peinado , M. A. , Watkins , D. N. , Issa , J. P. , Sidransky , D. , Baylin , S. B. and Herman , J. G.Inactivation of the DNA repair gene O6‐methylguanine‐DNA methyltransferase by promoter hypermethylation is associated with G to A mutations in K‐ras in colorectal tumorigenesis . Cancer Res. , 60 , 2368 – 2371 ( 2000. ). [PubMed] [Google Scholar]
  • 15. ) Simpson , D. J. , Hibberts , N. A. , McNicol , A. M. , Clayton , R. N. and Farrell , W. E.Loss of pRb expression in pituitary adenomas is associated with methylation of the RB1 CpG island . Cancer Res. , 60 , 1211 – 1216 ( 2000. ). [PubMed] [Google Scholar]
  • 16. ) Nakamura , M. , Sakaki , T. , Hashimoto , H. , Nakase , H. , Ishida , E. , Shimada , K. and Konishi , N.Frequent alterations of the p14ARF and p161NK4a genes in primary central nervous system lymphomas . Cancer Res. , 61 , 6335 – 6339 ( 2001. ). [PubMed] [Google Scholar]
  • 17. ) Nakamura , M. , Watanabe , T. , Yonekawa , Y. , Kleihues , P. and Ohgaki , H.Promoter methylation of the DNA repair gene MGMT in astrocytomas is frequently associated with G:C→A:T mutations of the TP53 tumor suppressor gene . Carcinogenesis , 22 , 1715 – 1719 ( 2001. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Xing , E. P. , Nie , Y. , Song , Y. , Yang , G. Y. , Cai , Y. C. , Wang , L. D. and Yang , C. S.Mechanisms of inactivation of p14ARF, p15INK4b, and P16INK4a genes in human esophageal squamous cell carcinoma . Clin. Cancer Res. , 5 , 2704 – 2713 ( 1999. ). [PubMed] [Google Scholar]
  • 19. ) Ishii , N. , Maier , D. , Merlo , A. , Tada , M. , Sawamura , Y. , Diserens , A. C. and Van Meir , E. G.Frequent co‐alterations of TP53, p16/CDKN2A, p14ARF, PTEN tumor suppressor genes in human glioma cell lines . Brain Pathol. , 9 , 469 – 479 ( 1999. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20. ) Konishi , N. , Hiasa , Y. , Hayashi , I. , Matsuda , H. , Tsuzuki , T. , Ming , T. , Kitahori , Y. , Shiraishi , T. , Yatani , R. and Shimazaki , J.p53 mutations occur in clinical, but not latent, human prostate carcinoma . Jpn. J. Cancer Res. , 86 , 57 – 63 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. ) Konishi , N. , Hiasa , Y. , Tsuzuki , T. , Tao , M. , Enomoto , T. and Miller , G. J.Comparison of ras activation in prostate carcinoma in Japanese and American men . Prostate , 30 , 53 – 57 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Konishi , N. , Hiasa , Y. , Tsuzuki , T. , Matsuda , H. , Tao , M. , Nakamura , M. , Naitho , H. , Kitahori , Y. , Shiraishi , T. , Yatani , R. , Shimazaki , J. and Lin , J.‐C.Detection of RB, p16/CDKN2 and p15INK4B gene alterations with immunohis‐tochemical studies in human prostate carcinomas . Int. J. Oncol. , 8 , 107 – 112 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 23. ) Konishi , N. , Cho , M. , Yamamoto , K. and Hiasa , Y.Genetic changes in prostate cancer . Pathol. Int. , 47 , 735 – 747 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 24. ) Esteller , M. , Cordon‐Cardo , C. , Corn , P. G. , Meltzer , S. J. , Pohar , K. S. , Watkins , D. N. , Capella , G. , Peinado , M. A. , Matias‐Guiu , X. , Prat , J. , Baylin , S. B. and Herman , J. G.p14ARF silencing by promoter hypermethylation mediates abnormal intracellular localization of MDM2 . Cancer Res. , 61 , 2816 – 2821 ( 2001. ). [PubMed] [Google Scholar]
  • 25. ) Baur , A. S. , Shaw , P. , Burri , N. , Delacretaz , R , Bosnian , F. T. and Chaubert , P.Frequent methylation silencing of p151NK4b (MTS2) and p161NK4a (MTS1) in B‐cell and T‐cell lymphomas . Blood , 94 , 1773 – 1781 ( 1999. ). [PubMed] [Google Scholar]
  • 26. ) Esteller , M. , Corn , P. G. , Baylin , S. B. and Herman , J. G.A gene hypermethylation profile of human cancer . Cancer Res. , 61 , 3225 – 3229 ( 2001. ). [PubMed] [Google Scholar]
  • 27. ) Kamijo , T. , Weber , J. D. , Zambetti , G. , Zindy , F. , Roussel , M. F. and Sherr , C. J.Functional and physical interactions of the ARF tumor suppressor with p53 and Mdm2 . Proc. Nad. Acad. Sci. USA , 95 , 8292 – 8297 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28. ) Pomerantz , J. , Schreiber‐Agus , N. , Liegeois , N. J. , Silverman , A. , Alland , L. , Chin , L. , Poles , J. , Chen , K. , Orlow , L , Lee , H. W. , Cordon‐Cardo , C. and De Pinho , R. A.The Ink4a tumor suppressor gene product, p19ARF, interacts with MDM2 and neutralizes MDM2′s inhibition of p53 . Cell , 92 , 713 – 723 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 29. ) Nguyen , T. T. , Nguyen , C. T. , Gonzales , F. A. , Nichols , P. W. , Yu , M. C. and Jones , P. A.Analysis of cyclin‐dependent kinase inhibitor expression and methylation patterns in human prostate cancers . Prostate , 43 , 233 – 242 ( 2000. ). [DOI] [PubMed] [Google Scholar]
  • 30. ) Konishi , N. , Nakamura , M. , Kishi , M. , Nishimine , M. , Ishida , E. and Shimada , K.Heterogeneous methylation and deletion patterns of the INK4a/ARF locus within prostate carcinomas . Am. J. Pathol. , 160 , 1207 – 1214 ( 2002. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31. ) Aaltomaa , S. , Lipponen , P. , Eskelinen , M. , Ala‐Opas , M. and Kosma , V. M.Prognostic value and expression of p21waf1/ciplprotein in prostate cancer . Prostate , 39 , 8 – 15 ( 1999. ). [DOI] [PubMed] [Google Scholar]
  • 32. ) Cheng , L. , Lloyd , R. V. , Weaver , A. L. , Pisansky , T. M. , Cheville , J. C. , Ramnani , D. M. , Leibovich , B. C. , Blute , M. L. , Zincke , H. and Bostwick , D. G.The cell cycle inhibitors p21WAF1 and p27KIP1 are associated with survival in patients treated by salvage prostatectomy after radiation therapy . Clin. Cancer Res. , 6 , 1896 – 1899 ( 2000. ). [PubMed] [Google Scholar]
  • 33. ) Cairns , P. , Okami , K. , Halachmi , S. , Halachmi , N. , Esteller , M. , Herman , J. G. , Jen , J. , Isaacs , W. B. , Bova , G. S. and Sidransky , D.Frequent inactivation of PTEN/MMAC1 in primary prostate cancer . Cancer Res. , 57 , 4997 – 5000 ( 1997. ). [PubMed] [Google Scholar]
  • 34. ) Whang , Y. E. , Wu , X. , Suzuki , H. , Reiter , R. E. , Tran , C. , Vessella , R. L. , Said , J. W. , Isaacs , W. B. and Sawyers , C. L.Inactivation of the tumor suppressor PTEN/MMACl in advanced human prostate cancer through loss of expression . Proc. Natl. Acad. Sci. USA , 95 , 5246 – 5250 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35. ) Lee , W. H. , Morton , R. A. , Epstein , J. I , Brooks , J. D. , Campbell , P. A. , Bova , G. S. , Hsieh , W. S. , Isaacs , W. B. and Nelson , W. G.Cytidine methylation of regulatory sequences near the pi‐class glutathione S‐transferase gene accompanies human prostatic carcinogenesis . Proc. Natl. Acad. Sci. USA , 91 , 11733 – 11737 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36. ) Santourlidis , S. , Florl , A. , Ackermann , R. , Wirtz , H. C. and Schulz , W. A.High frequency of alterations in DNA methylation in adenocarcinoma of the prostate . Prostate , 39 , 166 – 174 ( 1999. ). [DOI] [PubMed] [Google Scholar]
  • 37. ) Brooks , J. D. , Weinstein , M. , Lin , X. , Sun , Y. , Pin , S. S. , Bova , G. S. , Epstein , J. I , Isaacs , W. B. and Nelson , W. G.CG island methylation changes near the GSTP1 gene in prostatic intraepithelial neoplasia . Cancer Epidemiol. Biomarkers Prev. , 7 , 531 – 536 ( 1998. ). [PubMed] [Google Scholar]
  • 38. ) Esteller , M. , Corn , P. G. , Urena , J. M. , Gabrielson , E. , Baylin , S. B. and Herman , J. G.Inactivation of glutathione S‐transferase P1 gene by promoter hypermethylation in human neoplasia . Cancer Res. , 58 , 4515 – 4518 ( 1998. ). [PubMed] [Google Scholar]
  • 39. ) Aquilina , G. , Biondo , R. , Dogliotti , E. , Meuth , M. and Bignami , M.Expression of the endogenous O6‐methylguanine‐DNA‐methyltransferase protects Chinese hamster ovary cells from spontaneous G:C to A:T transitions . Cancer Res. , 52 , 6471 – 6475 ( 1992. ). [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES