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Archives of Pharmacal Research

, Volume 29, Issue 2, pp 152–158 | Cite as

Overexpression of p73 enhances cisplatin-induced apoptosis in HeLa cells

  • Keun-Cheol Kim
  • Chul-Soo Jung
  • Kyung-Hee Choi
Articles Drug Design

Abstract

To examine a possible synergistic role for p73 and cisplatin (cis-diamminedichloroplatinum II) in HeLa cells with a nonfunctional p53 protein, we established stable HeLa/p73 clones using a tetracycline inducible eukaryotic expression vector. The HeLa/p73 clones were not characterized by changes in growth or morphology. Cell death analysis, however, indicated a greater sensitivity to cisplatin in the p73-overexpressed HeLa cells than determined for the non-induced HeLa cells. This increased sensitivity seems to affect an induction of a sub-G1 population as assessed from flow cytometry analysis. The increased sub-G1 population may, in turn, result from a reduction of cyclin D1 and B1 expression by cisplatin in the presence of p73. Hoechest staining indicated an increased number of dead cells in the p73-induced cells compared to the non-induced cells. Poly ADP-ribose polymerase (PARP) cleavage was shown to be distinct in the p73-overexpressed cells compared to non-induced cells, which suggests that p73 modulates the cisplatin-induced apoptosis. Therefore, a synergistic effect of p73 and cisplatin to induce apoptosis could lead to new treatment for some types of human cancers.

Key words

Cisplatin p73 Drug sensitivity Cell death Apoptosis 

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References

  1. Agami, R., Blandino, G., Oren, M., and Shaul, Y., Interaction of c-Abl and p73a and their collaboration to induce apoptosis.Nature, 399, 809–813 (1999).PubMedCrossRefGoogle Scholar
  2. Bunz, F., Hwang, P. M., Torrance, C., Waldman, T., Zhang, Y., Dillehay, L., Williams, J., Lengauer, C., Kinzler, K., and Vogelstein, B., Disruption of p53 in human cancer cells alters the responses to therapeutic agents.J. Clin. Invest., 104, 263–269 (1999).PubMedCrossRefGoogle Scholar
  3. Cascallo, M., Calbo, J., Gelpi, J. L., and Mazo, A., Modulation of drug cytotoxicity by reintroduction of wild type p53 gene (Ad5CMV-p53) in human pancreatic cancer.Cancer Gene Ther., 7, 545–556 (2000).PubMedCrossRefGoogle Scholar
  4. Duan, L., Aoyagi, M., Tamaki, M., Yoshino, Y., Morimoto, T., Wakimoto, H., Nagasaka, Y., Hirakawa, K., Ohno, K., and Yamamoto, K., Impairment of both apoptotic and cytoprotective signalings in glioma cells resistant to the combined use of cisplatin and tumor necrosis factor alpha.Clin. Cancer Res., 10, 234–243 (2004).PubMedCrossRefGoogle Scholar
  5. Dubrez, L., Coll, J. L., Hurbin, A., Solary, E., and Favrot, M. C., Caffeine sensitizes human H358 cell line to p53-mediated apoptosis by inducing mitochondrial translocation and conformational change of BAX protein.J. Biol. Chem., 276, 38980–38987 (2001).PubMedCrossRefGoogle Scholar
  6. Dunkern, T. R., Fritz, G., and Kaina, B., Cisplatin-induced apoptosis in 43-3B and 27-1 cells defective in nucleotide excision repair.Mutat. Res., 486, 249–258 (2001).PubMedGoogle Scholar
  7. Fang, L., Igarashi, M., Leung, J., Sugrue, M., Lee, S., and Aaronson, S., p21 Waf2/Cip1/Sdi1 induces permanent growth arrest with markers of replicative senescence in tumor cells lacking functional p53.Oncogene, 18, 2789–2797 (1999).PubMedCrossRefGoogle Scholar
  8. Feng, L., Achanta, G., Pelicano, H., Zhang, W., Plunkett, W., and Huang, P., Role of p53 in cellular response to anticancer nucleoside analog-induced DNA damage.Int. J. Mol. Med., 5, 597–604 (2000).PubMedGoogle Scholar
  9. Gong, J. G., Costanzo, A., Yang, H.Q., Melino, G., Kaelin, W. G. Jr., Levrero, M., and Wang, J. Y., The tyrosine kinase c-Abl regulates p73 in apoptotic response to cisplatin-induced DNA damage.Nature, 399, 806–809 (1999).PubMedCrossRefGoogle Scholar
  10. Hannun, Y. A., Apoptosis and the dilemma of cancer chemotherapy.Blood, 89, 1845–1853 (1997).PubMedGoogle Scholar
  11. Husain, A., Rosales, N., Schwartz, G. K., and Spriggs, D. R., Lisophylline sensitizes p53 mutant human ovarian carcinima cells to the cytotoxic effects ofcis-diamminedichloroplatinum (II).Gynecol. oncol., 70, 17–22 (1998).PubMedCrossRefGoogle Scholar
  12. Jost, C. A., Marin, M. C., and Kaelin, W. G. Jr., p73 is a human p53-related protein that can induce apoptosis.Nature, 389, 191–194 (1997).PubMedCrossRefGoogle Scholar
  13. Kaghad, M., Bonnet, H., Yang, A., Creancier, L., Biscan, J. C., Valent, A., Minty, A., Chalon, P., Lelias, J. M., Dumont, X., Ferrara, P., McKeon, F., and Caput, D., Monoallelically expressed gene related to p53 at 1p36, a region frequently deleted in neuroblastoma and other human cancers.Cell, 90, 809–819 (1997).PubMedCrossRefGoogle Scholar
  14. Kang, K. H., Lee, J. H., Kim, K. C., Ham, S. W., Kim, M. Y., and Choi, K. H., Induction of p73beta by a naphthoquinone analog is mediated by E2F-1 and triggers apoptosis in HeLa cells.FEBS Lett., 522, 161–167 (2002).PubMedCrossRefGoogle Scholar
  15. Kang, K. H., Kim, W. H., and Choi, K. H., p21 promotes ceramide-induced apoptosis and antagonizes the antideath effect of bcl-2 in human hepatocarcinoma cells.Exp. Cell Res., 253, 403–412 (1999).PubMedCrossRefGoogle Scholar
  16. Kessis, T. D., Slebos, R. J., Nelson, W. G., Kastan, M. B., Plunkett, B. S., Han, S. M., Lorincz, A. T., Hedrick, L., and Cho, K. R., Human papillomavirus 16 E6 expression disrupts the p53-mediated cellular response to DNA damage.Proc. Natl. Acad. Sci. U.S.A., 90, 3988–3992 (1993).PubMedCrossRefGoogle Scholar
  17. Kim, K. C., Kim, T. S., Kang, K. H., and Choi, K. H., Amphiphysin IIb-1, a novel splicing variant of amphiphysin II, regulates p73beta function through protein-protein interactions.Oncogene, 20, 6689–6699 (2001).PubMedCrossRefGoogle Scholar
  18. Kim, R., Emi, M., Tanabe, K., and Toge, T., Therapeutic potential of antisense Bcl-2 as a chemosensitizer for cancer therapy.Cancer, 101, 2491–2502 (2004).PubMedCrossRefGoogle Scholar
  19. Lee, R. H., Song, J. M., Park, M. Y., Kang, S. K., Kim, Y. K., and Jung, J. S., Cisplatin-induced apoptosis by translocation of endogenous Bax in mouse collecting duct cells.Biochem. Pharmacol., 62, 1013–1023 (2001).PubMedCrossRefGoogle Scholar
  20. Minagawa, Y., Kigawa, J., Itamochi, H., Kanamori, Y., Shimada, M., Takahashi, M., and Terakawa, N., Cisplatin-resistant HeLa cells are resistant to apoptosis via p53-dependent and-independent pathways.Jpn. J. Cancer Res., 90, 1373–1379 (1999).PubMedGoogle Scholar
  21. Poppenborg, H., Munstermann, G., Knupfer, M. M., Hotfilder, M., and Wolff, J. E. A., C6 cells cross-resistant to cisplatin and radiation.Anticancer Res., 17, 2073–2078 (1997).PubMedGoogle Scholar
  22. Puig, P., Capodieci, P., Drobnjak M., Verbel, D., Prives, C., Cordon-Cardo, C., and Di Como, C. J., p73 Expression in human normal and tumor tissues: loss of p73α expression is associated with tumor progression in bladder cancerClin. Cancer Res., 9, 5642–5651 (2003).PubMedGoogle Scholar
  23. Qin, L. F. and Ng, I. O., Exogenous expression of p21(WAF1/CIP1) exerts cell growth inhibition and enhances sensitivity to cisplatin in hepatoma cells.Cancer Lett., 172, 7–15 (2001).PubMedCrossRefGoogle Scholar
  24. Radosevic, N., Delmer, A., Tang, R., Marie, J. P., and Ajchenbaum-Cymbalista, F., Cell cycle regulatory protein expression in fresh acute myeloid leukemia cells and after drug exposure.Leukemia, 15, 559–566 (2001).PubMedCrossRefGoogle Scholar
  25. Sphyris, N., Morris, R. G., and Harrison, D. J., Induction of p21 and nuclear accumulation of TAp73alpha and c-abl during apoptosis of cisplatin-treated primary pancreatic acinar cells.Int. J. Oncol., 25, 1661–1670 (2004).PubMedGoogle Scholar
  26. Vaisman, A., Varchenko, M., Said, I., and Chaney, S. G., Cell cycle changes associated with formation of Pt-DNA adducts in human ovarian carcinoma cells with different cisplatin sensitivity.Cytometry, 27, 54–64 (1997).PubMedCrossRefGoogle Scholar
  27. Wei, Q., Pitx2a binds to human papillomavirus type 18 E6 protein and inhibits E6-mediated P53 degradation in HeLa cells.J. Biol. Chem., 280, 37790–37797 (2005).PubMedCrossRefGoogle Scholar
  28. Yuan, Z. M., Shioya, H., Ishiko, T., Sun, X., Gu, J., Huang, Y. Y., Lu, H., Kharbanda, S., Weichselbaum, R., and Kufe, D., p73 is regulated by tyrosine kinase c-abl in the apoptotic response to DNA damage.Nature, 399, 814–817 (1999).PubMedCrossRefGoogle Scholar

Copyright information

© The Pharmaceutical Society of Korea 2006

Authors and Affiliations

  1. 1.Division of Life Sciences, College of Natural SciencesKangwon National UniversityChuncheonKorea
  2. 2.Department of Life Science, College of Natural SciencesChung-Ang UniversitySeoulKorea

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