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  • Original Paper
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Reconstitution of hTERT restores tumorigenicity in melanoma-derived c-Myc low-expressing clones

Oncogene volume 21, pages 3011–3019 (2002)Cite this article

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.

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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

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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.

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Authors and Affiliations

  1. Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute, Rome, Italy

    Annamaria Biroccio, Sarah Amodei, Barbara Benassi, Marco Scarsella, Donatella Del Bufalo, Carlo Leonetti & Gabriella Zupi

  2. Clinical and Pathology Laboratories, Regina Elena Cancer Institute, Rome, Italy

    Anna Cianciulli & Marcella Mottolese

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  1. Annamaria Biroccio
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  2. Sarah Amodei
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Correspondence to Gabriella Zupi.

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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

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