Abstract
Gefitinib is an anti-cancer drug that selectively inhibits epithelial growth factor receptor (EGFR) tyrosine kinase activity and induces apoptosis in many cancer cells. Cancer cells are often protected from apoptotic cell death by telomerase, however the gefitinib-induced telomerase inhibition remains unknown. Here we investigated the effects of gefitinib on telomerase activity in two different breast cancer lines, MCF-7 (low expression of EGFR) and MDA-MB-231 (high expression of EGFR). We observed the inhibition of EGFR phosphorylation that occurred only MDA-MB-231 cells cultured in media containing 10% FBS. Direct cytotoxicity was observed in MDA-MB-231 cells, but not MCF-7 cells when treated with concentrations of gefitinib ranging from 15 to 20 µM. This cytotoxicity was associated with decreased telomerase activity and downregulation of the telomerase subunit, hTERT. c-Myc has previously been shown to activate telomerase activity through transcriptional regulation of hTERT. A decrease in c-myc expression and DNA-binding activity following treatment with gefitinib was observed exclusively in MDA-MB-231 cells. We also demonstrated that gefitinib downregulates the activation of Akt and subsequent hTERT phosphorylation and translocation into the nucleus in MDA-MB-231 cells. These results indicate that gefitinib induces loss of telomerase activity through dephosphorylation of EGFR in MDA-MB-231 cells.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Anderson, N. G., Ahmad, T., Chan, K., Dobson, R., and Bundred, N. J., ZD1839 (Iressa), a novel epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, potently inhibits the growth of EGFR-positive cancer cell lines with or without erbB2 overexpression. Int. J. Cancer, 94, 774–782 (2001).
Chang, G. C., Hsu, S. L., Tsai, J. R., Liang, F. P., Lin, S. Y., Sheu, G. T., and Chen, C. Y., Molecular mechanisms of ZD1839-induced G1-cell cycle arrest and apoptosis in human lung adenocarcinoma A549 cells. Biochem. Pharmacol., 68, 1453–1464 (2004).
Ciardiello, F., Caputo, R., Borriello, G., Del Bufalo, D., Biroccio, A., Zupi, G., Bianco, A. R., and Tortora, G., ZD1839 (IRESSA), an EGFR-selective tyrosine kinase inhibitor, enhances taxane activity in bcl-2 overexpressing, multidrug-resistant MCF-7 ADR human breast cancer cells. Int. J. Cancer, 98, 463–469 (2002).
Claassen, G. F. and Hann, S. R., Myc-mediated transformation: the repression connection. Oncogene, 18, 2925–2933 (1999).
Cong, Y. S., Wen, J., and Bacchetti, S., The human telomerase catalytic subunit hTERT: organization of the gene and characterization of the promoter. Hum. Mol. Genet., 8, 137–142 (1999).
Feng, J., Funk, W. D., Wang, S. S., Weinrich, S. L., Avilion, A. A., Chiu, C. P., Adams, R. R., Chang, E., Yu, J., Le, S., West, M. D., Harley, C. B., Andrews, W. H., Greider, C. W., and Willeponteau B., The RNA component of human telomerase. Science, 269, 1236–1241 (1995).
Ghosh, U. and Bhattacharyya, N. P., Benzamide and 4-amino 1,8 naphthalimide treatment inhibit telomerase activity by down-regulating the expression of telomerase associated protein and inhibiting the poly(ADP-ribosyl)ation of telomerase reverse transcriptase in cultured cells. FEBS J., 272, 4237–4248 (2005).
Gilmore, A. P., Valentijn, A. J., Wang, P., Ranger, A. M., Bundred, N., O’Hare, M. J., Wakeling, A., Korsmeyer, S. J., and Streuli, C. H., Activation of BAD by therapeutic inhibition of epidermal growth factor receptor and transactivation by insulin-like growth factor receptor. J. Biol. Chem., 277, 27643–27650 (2002).
Greenberg, R. A., O’Hagan, R. C., Deng, H., Xiao, Q., Hann, S. R., Adams, R. R., Lichtsteiner, S., Chin, L., Morin, G. B., and DePinho, R. A., Telomerase reverse transcriptase gene is a direct target of c-Myc but is not functionally equivalent in cellular transformation. Oncogene, 18, 1219–1226 (1999).
Greider, C. W. and Blackburn, E. H., Identification of a specific telomere terminal transferase activity in Tetrahymena extracts. Cell, 43, 405–413 (1985).
Greider, C. W. and Blackburn, E. H., A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis. Nature, 337, 331–337 (1989).
Harrington, L., McPhail, T., Mar, V., Zhou, W., Oulton, R., Bass, M. B., Arruda, I., and Robinson, M. O., A mammalian telomerase-associated protein. Science, 275, 973–977 (1997).
Holt, S. E., Aisner, D. L., Baur, J., Tesmer, V. M., Dy, M., Ouellette, M., Trager, J. B., Morin, G. B., Toft, D. O., Shay, J. W., Wright, W. E., and White, M. A., Functional requirement of p23 and Hsp90 in telomerase complexes. Genes Dev., 13, 817–826 (1999).
Jagadeesh, S., Kyo, S., and Banerjee, P. P., Genistein represses telomerase activity via both transcriptional and posttranslational mechanisms in human prostate cancer cells. Cancer Res., 66, 2107–2115 (2006).
Janmaat, M. L., Kruyt, F. A., Rodriguez, J. A., and Giaccone, G., Response to epidermal growth factor receptor inhibitors in non-small cell lung cancer cells: limited antiproliferative effects and absence of apoptosis associated with persistent activity of extracellular signal-regulated kinase or Akt kinase pathways. Clin. Cancer Res., 9, 2316–2326 (2003).
Kang, S. S., Kwon, T., Kwon, D. Y., and Do, S. I., Akt protein kinase enhances human telomerase activity through phosphorylation of telomerase reverse transcriptase subunit. J. Biol. Chem., 274, 13085–13090 (1999).
Li, H., Zhao, L., Funder, J. W., and Liu, J. P., Protein phosphatase 2A inhibits nuclear telomerase activity in human breast cancer cells. J. Biol. Chem., 272, 16729–16732 (1997).
Li, H., Zhao, L., Yang, Z., Funder, J. W., and Liu, J. P., Telomerase is controlled by protein kinase C alpha in human breast cancer cells. J. Biol. Chem., 273, 33436–33442 (1998).
Li, H., Lee, T. H., and Avraham, H., A novel tricomplex of BRCA1, Nmi, and c-Myc inhibits c-Myc-induced human telomerase reverse transcriptase gene (hTERT) promoter activity in breast cancer. J. Biol. Chem., 277, 20965–20973 (2002).
Liu, J. P., Studies of the molecular mechanisms in the regulation of telomerase activity. FASEB J., 13, 2091–2104 (1999).
Liu, K., Hodes, R. J., and Weng, N. P., Cutting edge: telomerase activation in human T lymphocytes does not require increase in telomerase reverse transcriptase (hTERT) protein but is associated with hTERT phosphorylation and nuclear translocation. J. Immunol., 166, 4826–4830 (2001).
Magne, N., Fischel, J. L., Tiffon, C., Formento, P., Dubreuil, A., Renee, N., Formento, J. L., Francoual, M., Ciccolini, J., Etienne, M. C., and Milano, G., Molecular mechanisms underlying the interaction between ZD1839 (’Iressa’) and cisplatin/5-fluorouracil. Br. J. Cancer, 89, 585–592 (2003).
Meyerson, M., Counter, C. M., Eaton, E. N., Ellisen, L. W., Steiner, P., Caddle, S. D., Ziaugra, L., Beijersbergen, R. L., Davidoff, M. J., Liu, Q., Bacchetti, S., Haber, D. A., and Weinberg R. A., hEST2, the putative human telomerase catalytic subunit gene, is up-regulated in tumor cells and during immortalization. Cell, 90, 785–795 (1997).
Moasser, M. M., Basso, A., Averbuch, S. D., and Rosen, N., The tyrosine kinase inhibitor ZD1839 (“Iressa”) inhibits HER2-driven signaling and suppresses the growth of HER2-overexpressing tumor cells. Cancer Res., 61, 7184–7188 (2001).
Moulder, S. L., Yakes, F. M., Muthuswamy, S. K., Bianco, R., Simpson, J. F., and Arteaga, C. L., Epidermal growth factor receptor (HER1) tyrosine kinase inhibitor ZD1839 (Iressa) inhibits HER2/neu (erbB2)-overexpressing breast cancer cells in vitro and in vivo. Cancer Res., 61, 8887–8895 (2001).
Nakamura, T. M., Morin, G. B., Chapman, K. B., Weinrich, S. L., Andrews, W. H., Lingner, J., Harley, C. B., and Cech, T. R., Telomerase catalytic subunit homologs from fission yeast and human. Science, 277, 955–959 (1997).
Nicholson, R. I., Gee, J. M., and Harper, M. E., EGFR and cancer prognosis. Eur. J. Cancer, 37, S9–15 (2001).
Ouchi, H., Ishiguro, H., Ikeda, N., Hori, M., Kubota, Y., and Uemura, H., Genistein induces cell growth inhibition in prostate cancer through the suppression of telomerase activity. Int. J. Urol., 12, 73–80 (2005).
Rho, J. K., Choi, Y. J., Ryoo, B. Y., Na, I. I., Yang, S. H., Kim, C. H., and Lee, J. C., p53 enhances gefitinib-induced growth inhibition and apoptosis by regulation of Fas in non-small cell lung cancer. Cancer Res., 67, 1163–1169 (2007).
Sebastian, S., Grammatica, L., and Paradiso, A., Telomeres, telomerase and oral cancer (Review). Int. J. Oncol., 27, 1583–1596 (2005).
Sedivy, J. M., Can ends justify the means?: telomeres and the mechanisms of replicative senescence and immortalization in mammalian cells. Proc. Natl. Acad. Sci. USA, 95, 9078–9081 (1998).
Suenaga, M., Yamaguchi, A., Soda, H., Orihara, K., Tokito, Y., Sakaki, Y., Umehara, M., Terashi, K., Kawamata, N., Oka, M., Kohno, S., and Tei, C., Antiproliferative effects of gefitinib are associated with suppression of E2F-1 expression and telomerase activity. Anticancer Res., 26, 3387–3391 (2006).
Ulaner, G. A., Hu, J. F., Vu, T. H., Giudice, L. C., and Hoffman, A. R., Telomerase activity in human development is regulated by human telomerase reverse transcriptase (hTERT) transcription and by alternate splicing of hTERT transcripts. Cancer Res., 58, 4168–4172 (1998).
Wakeling, A. E., Guy, S. P., Woodburn, J. R., Ashton, S. E., Curry, B. J., Barker, A. J., and Gibson, K. H., ZD1839 (Iressa): an orally active inhibitor of epidermal growth factor signaling with potential for cancer therapy. Cancer Res., 62, 5749–5754 (2002).
Wang, J., Xie, L. Y., Allan, S., Beach, D., and Hannonm, G. J., Myc activates telomerase. Genes Dev., 12, 1769–1774 (1998).
Wu, K. J., Grandori, C., Amacker, M., Simon-Vermot, N., Polack, A., Lingner, J., and Dalla-Favera, R., Direct activation of TERT transcription by c-MYC. Nat. Genet., 21, 220–224 (1999).
Yan, C. H., Chen, X. G., Li, Y., and Han, R., Effects of genistein, a soybean-derived isoflavone, on proliferation and differentiation of B16-BL6 mouse melanoma cells. J. Asian Nat. Prod. Res., 1, 285–299 (1999).
Yeo, M., Rha, S. Y., Jeung, H. C., Hu, S. X., Yang, S. H., Kim, Y. S., An, S. W., and Chung, H. C., Attenuation of telomerase activity by hammerhead ribozyme targeting human telomerase RNA induces growth retardation and apoptosis in human breast tumor cells. Int. J. Cancer, 114, 484–489 (2005).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Moon, DO., Kim, MO., Heo, MS. et al. Gefitinib induces apoptosis and decreases telomerase activity in MDA-MB-231 human breast cancer cells. Arch. Pharm. Res. 32, 1351–1360 (2009). https://doi.org/10.1007/s12272-009-2002-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12272-009-2002-7