Skip to main content
Log in

Increased hTR Expression During Transition from Adenoma to Carcinoma Is Not Associated with Promoter Methylation

Digestive Diseases and Sciences Aims and scope Submit manuscript

Abstract

Human telomerase RNA component (hTR) expression, which increases in the majority of cancer cells with an acquisition of telomerase activity, was concomitantly evaluated with methylation status and human telomerase reverse transcriptase (hTERT) expression in colorectal cancers and precursor lesions. hTR and hTERT expressions were detected by in situ hybridization and reverse transcription following polymerase chain reaction, respectively, in 15 colonic adenomas, 19 sporadic colonic cancers at various histological stages, and 3 normal colonic mucosa samples. The methylationstatus of hTR was evaluated by methylation-specific polymerase chain reaction following restriction endonuclease digestion and direct sequencing. hTERT expression was detected in 16 of 19 cancers. hTR expression was detected in all cancers including two cases of intramucosal carcinoma. No hTR signals were detected in the normal epithelium or in the adenomas with severe atypism. CpG dinucleotides in the 5'-untranslated region of hTR were completely unmethylated from −204 to −3 and mosaically methylated from −290 to −272, irrespective of the atypism. These results suggest that hTR expression is increased at the adenoma-to-carcinoma transition stage but is not always associated with hTERT expression. Hypomethylation of the hTR promoter region is not likely to be the main mechanism regulating hTR expression.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  1. Blackburn EH, Greider CW: Telomeres. New York: Cold Spring Harbor Laboratory Press, 1995

    Google Scholar 

  2. Harley CB, Futcher AB, Greider CW: Telomeres shorten during aging of human fibroblasts. Nature 345:458–460, 1990

    Google Scholar 

  3. Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PL, Coviello GM, Wright WE, Weinrich SL, Shay JW: Specific association of human telomerase activity with immortal cells and cancer. Science 266:2011–2015, 1994

    Google Scholar 

  4. Bodnar AG, Ouellette M, Frolkis M, Holt SE, Chiu CP, Morin GB, Harley CB, Shay JW, Lichtsteiner S, Wright WE: Extension of lifespan by introduction of telomerase into normal humans. Science 279:349–352, 1998

    Google Scholar 

  5. Greider CW, Blackburn EH: Identification of a specific telomere terminal transferase activity in Tetrahymena extracts. Cell 43:405–413, 1985

    Google Scholar 

  6. Feng J, Funk WD, Wang SS, Weinrich SL, Avilion AA, Chiu CP, Adams RR, Chang E, Allsopp RC, Yu J, Le S, West MD, Harley CB, Andrews WH, Greider CW, Villeponteau B: The RNA component of human telomerase. Science 269:1236–1241, 1995

    Google Scholar 

  7. Nakamura TM, Morin GB, Chapman KB, Weinrich SL, Andrews WH, Lingner J, Harley CB, Cech TR: Telomerase catalytic subunit homologs from fission yeast and human. Science 277:955–959, 1997

    Google Scholar 

  8. Nakayama J, Saito M, Nakamura H, Matsuura A, Ishikawa F: TLP1: A gene encoding a protein component of mammalian telomerase is a novel member of WD repeats family. Cell 88:875–884, 1997

    Google Scholar 

  9. Harrington L, McPhail T, Mar V, Zhou W, Oulton R, Bass MB, Arruda I, Robinson MO: A mammalian telomerase-associated protein. Science 275:973–977, 1997

    Google Scholar 

  10. Weinrich SL, Pruzan R, Ma L, Ouellette M, Tesmer VM, Holt SE, Bodnar AG, Lichtsteiner S, Kim NW, Trager JB, Taylor RD, Carlos R, Andrews WH, Wright WE, Shay JW, Harley CB, Morin GB: Reconstitution of human telomerase with the templateRNAcomponent hTR and the catalytic protein subunit hTERT. Nat Gene 17:498–502, 1997

    Google Scholar 

  11. Avilion AA, Piatyszekm MA, Gupta J, Shay JW, Bacchetti S, Greider CW: Human telomerase RNA and telomerase activity in immortal cell lines and tumor tissues. Cancer Res 56:645–650, 1996

    Google Scholar 

  12. Nakano K, Watney E, McDougall JK: Telomerase activity and expression of telomerase RNA component and telomerase catalytic subunit gene in cervical cancer. Am J Pathol 153:857–864, 1998

    Google Scholar 

  13. Takahashi S, Kitamoto M, Takaishi H, Aikata H, Kawakami Y, Nakanishi T, Shimamoto F, Tahara E, Tahara, H, Ide T, Kajiyama G: Expression of telomerase component genes in hepatocellular carcinomas. Eur J Cancer 36:496–502, 2000

    Google Scholar 

  14. Kosciolek BA, Kalantidis K, Tabler M, Rowley PT: Inhibition of telomerase activity in human cancer cells by RNA interference. Mol Cancer Ther 2:209–216, 2003

    Google Scholar 

  15. Vulliamy T, Marrone A, Szydlo R, Walne A, Mason PJ, Dokal I: Disease anticipation is associated with progressive telomere shortening in families with dyskeratosis congenita due to mutations in TERC. Nat Genet 36:447–449, 2004

    Google Scholar 

  16. Zhao JQ, Hoare SF, McFarlane R, Muir S, Parkinson EK, Black DM, Keith WN: Cloning and characterization of human and mouse telomerase RNA gene promoter sequences. Oncogene 16:1345–1350, 1998

    Google Scholar 

  17. Wick M, Zubov D, Hagen G: Genomic organization and promoter characterization of the gene encoding the human telomerase reverse transcriptase (hTERT). Gene 232:97–106, 1999

    Google Scholar 

  18. Razin A, Riggs AD: DNA methylation and gene function. Science 210:604–610, 1980

    Google Scholar 

  19. Pogribny IP, Pogribna M, Christman JK, James SJ: Single-site methylation within the p53 promoter region reduces gene expression in a reporter gene construct: Possible in vivo relevance during tumorigenesis. Cancer Res 60:588–594, 2000

    Google Scholar 

  20. Cunningham JM, Christensen ER, Tester DJ, Kim CY, Roche PC, Burgart LJ, Thibodeau SN: Hypermethylation of the hMLH1 promoter in colon cancer with microsatellite instability. Cancer Res 58:3455–3460, 1998

    Google Scholar 

  21. Feinberg AP, Vogelstein B: Hypomethylation distinguishes genes of some human cancers from their normal counterparts. Nature 301:89–92, 1983

    Google Scholar 

  22. Lin CH, Hsieh SY, Sheen IS, Lee WC, Chen TC, Shyu WC, Liaw YF: Genome-wide hypomethylation in hepatocellular carcinogenesis. Cancer Res 61:4238–4243, 2001

    Google Scholar 

  23. World Medical Association Declaration of Helsinki: Ethical principles for medical research involving human subjects. Bull WHO 79:373–374, 2001

    Google Scholar 

  24. Japanese Society for Cancer of the Colon and Rectum: Japanese Classification of Colorectal Carcinoma, 1st English ed. Tokyo, Kanehara & Co., 1997

    Google Scholar 

  25. Raff T, van der Giet M, Endemann D, Wiederholt T, Paul M: Design and testing of beta-actin primers for RT-PCR that do not co-amplify processed pseudogenes. Biotechniques 23:456–460, 1997

    Google Scholar 

  26. Hoare SF, Bryce LA, Wisman GB, Burns S, Going JJ, van der Zee AG, Keith WN: Lack of telomerase RNA gene hTERC expression in alternative lengthening of telomeres cells is associated with methylation of the hTERC promoter. Cancer Res 61:27–32, 2001

    Google Scholar 

  27. Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB: Methylation-specific PCR: A novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA 93:9821–9826, 1996

    Google Scholar 

  28. Yi X, Tesmer V, M Savre-Train I, Shay JW, Wright WE: Both transcriptional and posttranscriptional mechanisms regulate human telomerase template RNA levels. Mol Cell Biol 19:3989–3997, 1999

    Google Scholar 

  29. Jones PA: The DNA methylation paradox. Trends Genet 15:34–37, 1999

    Google Scholar 

  30. Kuniyasu H, Domen T, Hamamoto T, Yokozaki H, Yasui W, Tahara H, Tahara E: Expression of human telomerase RNA is an early event of stomach carcinogenesis. Jpn J Cancer Res 88:103–107, 1997

    Google Scholar 

  31. Heine B, Hummel M, Demel G, Stein H: Demonstration of constant upregulation of the telomerase RNA component in human gastric carcinomas using in situ hybridization. J Patho 185:139–144, 1998

    Google Scholar 

  32. Ogami M, Ikura Y, Nishiguchi S, Kuroki T, Ueda M, Sakurai M: Quantitative analysis and in situ localization of human telomerase RNA in chronic liver disease and hepatocellular carcinoma. Lab Invest 79:15–26, 1999

    Google Scholar 

  33. Poremba C, Heine B, Diallo R, Heinecke A, Wai D, Schaefer KL, Braun Y, Schuck A, Lanvers C, Bankfalvi A, Kneif S, Torhorst J, Zuber M, Kochli OR, Mross F, Dieterich H, Sauter G, Stein H, Fogt F, Boecker W: Telomerase as a prognostic marker in breast cancer: high-throughput tissue microarray analysis of hTERT and hTR. J Patho 198:181–189, 2002

    Google Scholar 

  34. Lee RG: Benign tumors of the colon. In Bockus Gastroenterology, 5th ed. WS Haubrich, F Schaffner, JE Berk (eds). Philadelphia, Saunders, 1995, pp 1715–1730

    Google Scholar 

  35. Owen DA: Flat adenoma, flat carcinoma, and de novo carcinoma of the colon. Cancer 77:3–6, 1996

    Google Scholar 

  36. Hermanek P, Gall FP: Early (microinvasive) colorectal carcinoma pathology, diagnosis, surgical treatment. Int J Colorectal Dis 1:7–84, 1986

    Google Scholar 

  37. Shinya H, Wolff WI: Morphology, anatomic distribution and cancer potential of colonic polyps: An analysis of 7000 polyps endoscopically removed. Ann Surg 190:679–683, 1979

    Google Scholar 

  38. Stolte M, Bethke B: Colorectal mini-de novo carcinoma: A reality in Germany too. Endoscopy 27:286–290, 1995

    Google Scholar 

  39. Shimoda T, Ikegami M, Fujisaki J, Matsui T, Aizawa S, Ishikawa E: Early colorectal carcinoma with special reference to its development de novo. Cancer 64:1138–1146, 1989

    Google Scholar 

  40. Hirata I, Tanaka M, Sugimoto K, Yoshizumi M, Umegaki E, Masaki H: Clinicopathological study on flat and depressed minute colorectal carcinomas. Dig Endosc 3:526–535, 1991

    Google Scholar 

  41. Iishi H, Kitamura S, Nakaizumi A, Tatsuta M, Otani T, Okuda S: Clinicopathological features and endoscopic diagnosis of superficial early adenocarcinomas of the large intestine. Dig Dis Sci 38:1333–1337, 1993

    Google Scholar 

  42. Schlemper RJ, Itabashi M, Kato Y, Lewin KJ, Riddell RH, Shimoda T, Sipponen P, Stolte M, Watanabe H: Differences in the diagnostic criteria used by Japanese and Western pathologists to diagnose colorectal carcinoma. Cancer 82:60–69, 1998

    Google Scholar 

  43. Lewin KJ, Riddell RH, Weinstein WM: Gastrointestinal pathology and its clinical implications. New York, Igaku-Shoin, 1992, pp 1256–1257

    Google Scholar 

  44. Jass JR, Sobin LH: Histological typing of intestinal tumours. In WHO International Histological Classification of Tumours, 2nd ed. Berlin, Springer-Verlag, 1989, pp 29–40

    Google Scholar 

  45. Suda T, Isokawa O, Aoyagi Y, Nomoto M, Tsukada K, Shimizu T, Suzuki Y, Naito A, Igarashi H, Yanagi M, Takahashi T, Asakura H: Quantitation of telomerase activity in hepatocellular carcinoma: A possible aid for a prediction of recurrent diseases in the remnant liver. Hepatology 27:402–406, 1998

    Google Scholar 

  46. Chang JT, Chen YL, Yang HT, Chen CY, Cheng AJ: Differential regulation of telomerase activity by six telomerase subunits. Eur J Biochem 269:3442–3450, 2002

    Google Scholar 

  47. Henson JD, Neumann AA, Yeager TR, Reddel RR: Alternative lengthening of telomeres in mammalian cells. Oncogene 21:598–610, 2002

    Google Scholar 

  48. Bryan TM, Englezou A, Dalla-Pozza L, Dunham MA, Reddel RR: Evidence for an alternative mechanism for maintaining telomere length in human tumors and tumor-derived cell lines. Nat Med 3:1271–1274, 1997

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nakamura, A., Suda, T., Honma, T. et al. Increased hTR Expression During Transition from Adenoma to Carcinoma Is Not Associated with Promoter Methylation. Dig Dis Sci 49, 1504–1512 (2004). https://doi.org/10.1023/B:DDAS.0000042256.89282.c4

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:DDAS.0000042256.89282.c4

Navigation