Abstract
c-Myc is involved in the control of telomerase activity through its ability to induce the expression of the catalytic subunit of the enzyme, the human telomerase reverse transcriptase (hTERT). Our aim was to study whether telomerase plays a critical role in c-Myc-dependent tumorigenicity of melanoma cells. By using M14-derived clones, expressing low levels of c-Myc, we demonstrated that the down-regulation of c-Myc reduced cell proliferation rate, cloning efficiency and tumorigenicity and increased the apoptotic rate. Decreased tumorigenic potential correlated with reduced hTERT gene expression, telomerase activity and telomere shortening. Introduction of wild-type hTERT into these cells increased their proliferation rate and partially re-established their tumorigenic potential, at early passages, even though the apoptotic rate of the population remained unaltered. After several in vitro passages, hTERT-mediated cell proliferation made the tumorigenic potential of the c-Myc low-expressing clones comparable to that of the M14 parental line. Over-expression of the mutant biologically inactive hTERT did not drive cells to proliferate. In conclusion, our results demonstrate that the reconstitution of high levels of telomerase activity reverses the low tumorigenicity due to low c-Myc expression.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Abbreviations
- hTERT:
-
human Telomerase Reverse Transcriptase
- hTERT-HA:
-
Hemagglutinin-tagged human Telomerase Reverse Transcriptase
- wt:
-
wild-type
- mt:
-
mutant
- BrdU:
-
Bromodeoxyuridine
- TUNEL:
-
Terminal deoxynucleotide Transferase [TdT]-Mediated dUTP Nick-End Labeling
- RT:
-
Reverse Transcriptase
- PCR:
-
Polymerase Chain Reaction, TRAP, Telomeric Repeat Amplification Protocol
- ITAS:
-
Internal TRAP Assay Standard
- TRF:
-
Terminal Restriction Fragment
- mAb:
-
monoclonal antibody
References
Autexier C, Greider CW . 1996 Trends Biochem. Sci. 21: 387–391
Bello-Fernandez C, Packham G, Cleveland JL . 1993 Proc. Natl. Acad. Sci. USA 90: 7804–7808
Biroccio A, Benassi B, Amodei S, Gabellini C, Del Bufalo D, Zupi G . 2001 Mol. Pharm. 60: 174–182
Bodnar AG, Ouellette M, Frolkis M, Holt SE, Chiu CP, Morin GB, Harley CB, Shay JW, Lichtsteiner S, Wright WE . 1998 Science 279: 349–352
Chadeneau C, Siegel P, Harley CB, Muller WJ, Bacchetti S . 1995 Oncogene 11: 893–898
Chin L, Artandi SE, Shen Q, Tam A, Lee S, Gottlieb GJ, Greider CW, DePinho RA . 1999 Cell 97: 527–538
Citro G, D'Agnano I, Leonetti C, Perini R, Bucci B, Zon G, Calabretta B, Zupi G . 1998 Cancer Res. 58: 283–289
Cole MD . 1986 Ann. Rev. Genet. 20: 361–384
Counter CM, Hahn WC, Wei W, Caddle SD, Beijerbergen RL, Lansdorp PM, Sedivy JM, Weinberg RA . 1998 Proc. Natl. Acad. Sci. USA 95: 14723–14728
D'Cruz CM, Gunther EJ, Boxer RB, Hartman JL, Sintasath L, Moody SE, Cox JD, Ha SI, Belka GK, Golant A, Cardiff RD, Chodosh LA . 2001 Nat. Med. 7: 235–239
Dalla-Favera R, Bregni M, Erikson J, Patherson D, Gallo RC, Croce CM . 1982 Proc. Natl. Acad. Sci. USA 79: 7824–7827
Eilers M, Schirm S, Bishop JM . 1991 EMBO J. 10: 133–141
Fujimoto K, Takahashi M . 1997 Biochem. Biophys. 241: 775–781
Galaktionov K, Chen X, Beach D . 1996 Nature 382: 511–517
Gonzalez-Suarez E, Samper E, Flores JM, Blasco MA . 2000 Nat. Genet. 26: 114–117
Gonzalez-Suarez E, Samper E, Ramirez A, Flores JM, Martin-Caballero J, Jorcano JL, Blasco MA . 2001 EMBO J. 20: 2619–2630
Grandori C, Eisenman RN . 1997 Trends Biochem. Sci. 22: 177–181
Grandori C, Mac J, Siebelt F, Ayer DE, Eisenman RN . 1996 EMBO J. 15: 4344–4357
Greenberg R, Allsopp RC, Chin L, Morin GB, De Pinho RA . 1998 Oncogene 16: 1723–1730
Greenberg RA, Chin L, Femino A, Lee KH, Gottlieb GJ, Singer RH, Greider CW, DePinho RA . 1999b Cell 97: 515–525
Greenberg RA, O'Hagan RC, Deng H, Xiao Q, Hann SR, Adams RR, Lichtsteiner S, Chin L, Morin GB, DePinho RA . 1999a Oncogene 18: 1219–1226
Greider CW . 1998 Proc. Natl. Acad. Sci. USA 95: 90–92
Hahn WC, Counter CM, Lundberg AS, Beijersbergen RL, Brooks MW, Weinberg RA . 1999a Nature 400: 464–468
Hahn WC, Stewart SA, Brooks MW, York SG, Eaton E, Kurachi A, Beijersbergen RL, Knoll JHM, Meyerson M, Weinberg RA . 1999b Nat. Med. 10: 1164–1170
Harley CB, Fucher AB, Greider CW . 1990 Nature 345: 458–460
Herbert BS, Pitts AE, Baker SI, Hamilton SE, Wright WE, Shay JW, Corey DR . 1999 Proc. Natl. Acad. Sci. USA 96: 14276–14281
Hiyama E, Hiyama K, Yokoyama T, Matsuura Y, Piatyszek MA, Shay JW . 1995 Nat. Med. 1: 249–255
Jiang XR, Jimenez G, Chang E, Frolkis M, Kusler B, Sage M, Beeche M, Bodnar AG, Wahl GM, Tisty TD, Chiu CP . 1999 Nat. Genet. 21: 111–114
Kraehn GM, Utikal J, Udart M, Greulich KM, Bezold G, Kaskel P, Leiter U, Peter RU . 2001 Br. J. Cancer 84: 72–79
Kyo S, Takakura M, Taira T, Kanaya T, Itoh H, Yutsudo M, Ariga H, Inoue M . 2000 Nucleic Acids Res. 28: 669–677
Land H, Parada LF, Weinberg RA . 1983 Nature 304: 596–602
Lee LA, Dolde C, Barrett J, Wu CS, Dang CV . 1996 J. Clin. Invest. 97: 1687–1695
Lee TC, Li L, Philipson L, Ziff EB . 1997 Proc. Natl. Acad. Sci. USA 94: 12886–12891
Leonetti C, Biroccio A, Candiloro A, Citro G, Fornari C, Mottolese M, Del Bufalo D, Zupi G . 1999 Clin. Cancer Res. 5: 2588–2595
Leonetti C, D'Agnano I, Lozupone F, Valentini A, Geiser T, Zon G, Calabretta B, Citro G, Zupi G . 1996 J. Natl. Cancer Inst. 88: 419–429
Littlewood TD, Hancock DC, Danieilan PS, Parker MG, Evan GI . 1995 Nucleic Acids Res. 23: 1686–1690
Meyerson M, Counter CM, Eaton EN, Ellisen LW, Steiner P, Caddle SD, Ziaugra L, Beiijersbeergen RL, Davidoff MJ, Liu Q, Bacchetti S, Haber DA, Weinberg RA . 1997 Cell 90: 785–795
Morales CP, Holt SE, Ouellette M, Kaur KJ, Yan Y, Wilson KS, White MA, Wright WE, Shay JW . 1999 Nat. Genet. 21: 115–118
Nakamura TM, Morin GB, Chapman KB, Weinrich SL, Andrews WH, Lingner J, Harley CB, Cech TR . 1997 Science 277: 955–959
Shay JW, Bacchetti S . 1997 Eur. J. Cancer 33: 787–791
Shim H, Dolde C, Lewis BC, Wu CS, Dang G, Jungmann RA, Dalla-Favera R, Dang CV . 1997 Proc. Natl. Acad. Sci. USA 94: 6658–6663
Sinn E, Muller W, Pattengale P, Tepler I, Wallace R, Leder P . 1987 Cell 49: 465–475
Vaziri H, Benchimol S . 1998 Oncogene 8: 279–282
Wang J, Hannon GJ, Beach DH . 2000 Nature 405: 755–756
Wang J, Ying Xie L, Allan S, Beach D, Hannon GJ . 1998 Genes Dev. 12: 1769–1774
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 . 1997 Nat. Genet. 17: 498–502
Wu KJ, Grandori C, Amacker M, Simon-Vermot N, Polack A, Lingner J, Dalla-Favera R . 1999 Nat. Genet. 21: 220–224
Zhang X, Mar V, Zhou W, Harrington L, Robinson M . 1999 Genes Dev. 8: 2388–2399
Acknowledgements
We thank Dr Robert A Weinberg for the hTERT, hTERT-HA and DN-hTERT cDNA and Dr Garry P Nolan for the Phoenix amphotropic cells. We are extremely thankful to Dr S Bacchetti for very useful suggestions for the work and for extensive comments on the manuscript and to Dr A Farsetti for critical reading of the paper. S Amodei and B Benassi are recipients of a fellowship from the Italian Foundation for Cancer Research (FIRC). This work was supported by grants from Italian Association for Cancer Research (AIRC), Ministero della Sanità and CNR-MURST.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Biroccio, A., Amodei, S., Benassi, B. et al. Reconstitution of hTERT restores tumorigenicity in melanoma-derived c-Myc low-expressing clones. Oncogene 21, 3011–3019 (2002). https://doi.org/10.1038/sj.onc.1205415
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1205415
Keywords
This article is cited by
-
PARP1 is activated at telomeres upon G4 stabilization: possible target for telomere-based therapy
Oncogene (2010)
-
Multiple interference of the human papillomavirus-16 E7 oncoprotein with the functional role of the metastasis suppressor Nm23-H1 protein
Journal of Bioenergetics and Biomembranes (2006)
-
Involvement of hTERT in apoptosis induced by interference with Bcl-2 expression and function
Cell Death & Differentiation (2005)
-
Telomerase activity in colorectal carcinoma and its correlation with expression of c-myc
Chinese Journal of Cancer Research (2005)
-
Telomerase activity, apoptosis and cell cycle progression in ataxia telangiectasia lymphocytes expressing TCL1
British Journal of Cancer (2003)