Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

The mdm2 proto-oncogene sensitizes human medullary thyroid carcinoma cells to ionizing radiation

Oncogene volume 21, pages 2376–2386 (2002)Cite this article

Abstract

We have analysed the radiation response of a human medullary thyroid carcinoma cell line (MTT), characterized by the absence of a functional p53 protein, and the consequences of MDM2 overexpression in this process. We show that the product of the mdm2 proto-oncogene is able to sensitize MTT cells to ionizing radiation. After radiation treatment, MTT cells display histograms consistent with a G2M arrest. MTT cells expressing MDM2 (MTT-mdm2) are unable to respond to DNA damage with G2M arrest, and display a high percentage of apoptosis. MTT-mdm2 cells show high levels of E2F-1 protein, suggesting that the induction of apoptosis observed upon MDM2 overexpression could be dependent on E2F-1. This observation is further supported with assays showing that E2F-1 binding to specific DNA sequences is enhanced in MTT-mdm2 cells. Likewise, transactivation of reporter constructs exclusively dependent on E2F-1 is also elevated after transfection with MDM2. This effect can be reverted by transient transfection with p19ARF. To link the expression of E2F-1 with the induction of apoptosis, we generated clonal cell lines overexpressing E2F-1. Transfection with E2F-1 results in a low number of outgrowing colonies with reduced proliferation rates, indicating that E2F-1 is deleterious for cell growth. This negative regulation correlates with an increase in the percentage of the cell population with DNA content below 2N, suggesting that E2F-1 promotes apoptosis. Finally, overexpression of E2F-1 sensitizes MTT cells to radiation exposure. We conclude that the effects observed by MDM2 overexpression could be mediated by E2F-1.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  • Albright N . 1987 Radiat. Res. 112: 331–340

  • Alkhalaf M, Ganguli G, Messaddeq N, Le Meur M, Wasylyk B . 1999 Oncogene 18: 1419–1434

  • Andrews NC, Faller DV . 1991 Nucleic Acid Res. 19: 2499–

  • Banerjee D, Schnieders B, Fu JZ, Adhikari D, Zhao SC, Bertino JR . 1998 Cancer Res. 58: 4292–4296

  • Blattner C, Sparks A, Lane D . 1999 Mol. Cell. Biol. 19: 3704–3713

  • Brown DR, Thomas CA, Deb SP . 1998 EMBO J. 17: 2513–2525

  • Cachilly-Snyder L, Yang-Feng T, Francke U, George DL . 1987 Somatic Cell. Mol. Genet. 13: 235–244

  • Carlomagno F, Salvatore D, Santoro M, de Franciscis V, Quadro L, Panariello L, Colantuoni V, Fusco A . 1995 Biochem. Biophys. Res. Commun. 207: 1022–1028

  • Chang EH, Pirollo KF, Zou ZQ, Cheung HY, Lawler EL, Garner R, White E, Bernstein WB, Fraumeni JW, Blattner WA . 1987 Science 237: 1036–1038

  • Chen CY, Oliner JD, Zhan Q, Fornace AJ, Vogelstein B, Kastan MB . 1994 Proc. Natl. Acad. Sci. USA 91: 2684–2688

  • Dilla T, Velasco JA, Medina DL, Gonzalez-Palacios JF, Santisteban P . 2000 Endocrinology 141: 420–429

  • Dong YB, Yang HL, Jane M, Liu TJ, Stilwell A, Atienza C, McMasters KM . 1999 Cancer 86: 2021–2033

  • El-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B . 1993 Cell 75: 817–825

  • Finlay CA . 1993 Mol. Cell. Biol. 13: 301–306

  • Ganguli G, Abecassis J, Wasylyk B . 2000 EMBO J. 19: 5135–5147

  • Giuffrida D, Gharib H . 1998 Ann. Oncol. 9: 695–701

  • Haupt Y, Maya R, Kazaz A, Oren M . 1997 Nature 387: 296–299

  • Helin K, Lees JA, Vidal M, Dyson N, Harlow E, Fattaey A . 1992 Cell 24: 337–350

  • Heshmati HM, Gharib H, Van Heerden JA, Sizemore GW . 1997 Am. J. Med. 103: 60–69

  • Hunt KK, Deng J, Liu TJ, Wilson-Heiner M, Swisher SG, Clayman G, Hung MC . 1997 Cancer Res. 57: 4722–4726

  • Hwang A, Muschel RJ . 1998 Radiat. Res. 150: S52–S59

  • Illiakis G . 1997 Seminars Oncol. 24: 602–615

  • Johnson DG . 2000 Mol. Carcinog. 27: 151–157

  • Kasid U, Pfeiffer A, Brennan T, Beckett M, Weichselbaum RR, Dritschilo A, Mark GE . 1989 Science 243: 1354–1356

  • Kelle DE, Zyndi F, Ashmun RA, Sherr JC . 1995 Cell 83: 993–1000

  • Kubbutat MHG, Jones SN, Vousden KH . 1997 Nature 387: 299–303

  • Lam EW, Watson RJ . 1993 EMBO J. 12: 2705–2713

  • Leong SS, Horoszewicz JS, Shimaoka K, Friedman M, Kawinski E, Song MJ, Zeigel R, Chu TM, Bayling SB, Mirand EA . 1981 Advances in thyroid neoplasia Andreoli M, Monaco F, Robbins J (eds) Rome, Field Educational Italia pp 95–108

    Google Scholar 

  • Lin WC, Lin FT, Nevins JR . 2001 Genes Dev. 15: 1833–1844

  • Liu TJ, Wang M, Breau LR, Henderson Y, El Naggar AK, Steck KD, Siccard MW, Clayman GL . 1999 Cancer Gene Ther. 6: 163–171

  • Loughran O, La Thange NB . 2000 Mol. Cell. Biol. 20: 2186–2197

  • Lozano G, Montes de Oca Luna R . 1998 Biochim. Biophys. Acta 1377: M55–M59

  • Martin K, Trouche D, Hagemeler C, Sorensen TS, La Thange NB, Kouzarides T . 1995 Nature 375: 691–694

  • Medina DL, Velasco JA, Santisteban P . 1999 Endocrinology 140: 87–95

  • Momand J, Zambetti GP . 1997 J. Cell. Biochem. 64: 343–352

  • Momand J, Zambetti GP, Olson DC, George D, Levine AJ . 1992 Cell 69: 1237–1245

  • Nelkin BD, De Bustos A, Mabry M, Baylin SB . 1989 J. Amer. Med. Ass. 261: 3130–3135

  • Ohtani K . 1999 Front. Biosci. 4: D793–D804

  • Oliner JD, Kinzler KW, Meltzer PS, George DL, Vogelstein B . 1992 Nature 358: 80–83

  • Oswald F, Lovec H, Moroy T, Lipp M . 1994 Oncogene 9: 2029–2036

  • Phillips AC, Ernst MK, Bates S, Rice NR, Vousden KH . 1999 Mol. Cell 4: 771–781

  • Pirollo KF, Tong YA, Villegas Z, Chen Y, Chang EH . 1993 Radiat. Res. 135: 234–243

  • Puxeddu E, Fagin JA . 2001 Endocrinol Metab Clin North Am. 30: 493–513

  • Pruschy M, Wirbelauer C, Glanzmann C, Bodis S, Krek W . 1999 Cell Growth Differ. 10: 141–146

  • Sawey MJ, Hoot TA, Burns FJ, Garte SJ . 1987 Mol. Cell. Biol. 7: 932–935

  • Varghese S, Jung M . 1998 Arch. Pharma. Res. 21: 640–644

  • Vecchio G, Santoro M . 2000 Clin. Chem. Lab. Med. 38: 113–116

  • Velasco JA, Acebron A, Zannini M, Martin-Perez J, Di Lauro R, Santisteban P . 1998 Endocrinology 139: 2796–2802

  • Velasco JA, Medina DL, Romero J, Mato ME, Santisteban P . 1997 Int. J. Cancer 73: 449–455

  • Zhan Q, Fan S, Bae I, Guillouf C, Liebermann DA, O'Connor PM, Fornace Jr AJ . 1994 Oncogene 9: 3743–3751

  • Zhang R, Wang H . 2000 Current Pharmac. Design 6: 393–416

  • Zhang Y, Xiong Y, Yarbrough G . 1998 Cell 92: 725–734

Download references

Acknowledgements

We thank Dr Vicente Notario from Georgetown University Medical Center for critical reading of the manuscript. Tatiana Dilla is recipient of a fellow from the Fondo de Investigaciones Sanitarias. The work is supported by grants from the Comunidad de Madrid (CAM 08.1/0025/1997), Dirección General de Ciencia y Tecnología (BMC-2001-2087) and Fundación Salud 2000 (Spain).

Author information

Author notes

  1. Juan A Velasco

    Present address: Lilly Research Laboratories, Avda. de la Industria 30, Alcobendas, 28018, Madrid, Spain

Authors and Affiliations

  1. Instituto de Investigaciones Biomédicas Alberto Sols, Consejo Superior de Investigaciones Científicas, Madrid, Spain

    Tatiana Dilla & Pilar Sanstisteban

  2. Departamento de Oncología Radioterápica, Hospital Puerta de Hierro, Madrid, Spain

    Jesús Romero

  3. Molecular Oncology Program, Centro Nacional de Investigaciones Oncológicas Carlos III, Madrid, Spain

    Juan A Velasco

Authors
  1. Tatiana Dilla
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jesús Romero
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pilar Sanstisteban
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Juan A Velasco
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pilar Sanstisteban.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dilla, T., Romero, J., Sanstisteban, P. et al. The mdm2 proto-oncogene sensitizes human medullary thyroid carcinoma cells to ionizing radiation. Oncogene 21, 2376–2386 (2002). https://doi.org/10.1038/sj.onc.1205307

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1205307

Keywords

This article is cited by

Search

Advanced search

Quick links