Influence of G2 arrest on the cytotoxicity of DNA topoisomerase inhibitors toward human carcinoma cells with different p53 status

• Autorzy: Bozko P. , Larsen A.K. , Raymond E. , Skladanowski A.
• Języki publikacji: EN

Abstrakty

EN

We here report the influence of the cell cycle abrogator UCN-01 on RKO human co­lon carcinoma cells differing in p53 status following exposure to two DNA damaging agents, the topoisomerase inhibitors etoposide and camptothecin. Cells were treated with the two drugs at the IC90 concentration for 24 h followed by post-incubation in drug-free medium. RKO cells expressing wild-type, functional p53 arrested the cell cy­cle progression in both the Gj and G2 phases of the cell cycle whereas the RKO/E6 cells, which lack functional p53, only arrested in the G2 phase. Growth-arrested cells did not resume proliferation even after prolonged incubation in drug-free medium (up to 96 h). To evaluate the importance of the cell cycle arrest on cellular survival, a non-toxic dose of UCN-01 (100 nM) was added to the growth-arrested cells. The addi­tion of UCN-01 was accompanied by mitotic entry as revealed by the appearance of condensed chromatin and the MPM-2 phosphoepitope, which is characteristic for mi- totic cells. G2 exit and mitotic transit was accompanied by a rapid activation of caspase-3 and apoptotic cell death. The influence of UCN-01 on the long-term cytotoxic effects of the two drugs was also determined. Unexpectedly, abrogation of the G2 arrest had no influence on the overall cytotoxicity of either drug. In contrast, addition of UCN-01 to cisplatin-treated RKO and RKO/E6 cells greatly increased the cytotoxic effects of the alkylating agent. These results strongly suggest that even prolonged cell cycle arrest in the G2 phase of the cell cycle is not necessarily coupled to efficient DNA repair and enhanced cellular survival as generally believed.

Słowa kluczowe

EN

carcinoma cell; cell cycle; DNA topoisomerase inhibitor; man; cytotoxicity

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2002
• Tom: 49
• Numer: 1
• Opis fizyczny: p.109-119,fig.

Twórcy

autor

Bozko P.

• Technical University of Gdansk, Gdansk, Poland

autor

Larsen A.K.

autor

Raymond E.

autor

Skladanowski A.

Bibliografia

• 1.Hartwell, L.H. & Weinert, T.A. (1989) Checkpoints: Controls that ensure the order of cell cycle events. Science. 246, 629-634.
• 2.Graves, P.R., Yu, L., Schwarz, J.K., Gales, J., Sausville, E.A., O'Connor, P.M. & Piwnica- Worms, H. (2000) The Chkl protein kinase and the Cdc25C regulatory pathways are targets of the anticancer agent UCN-01. J. Biol. Chem. 275, 5600-5605.
• 3.Monks, A., Harris, E.D., Vaigro-Wolff, A., Hose, C.D., Connelly, J.W. & Sausville, E.A. (2000) UCN-01 enhances the in vitro toxicity of clinical agents in human tumor cell lines. Invest. New Drugs 18, 95-107.
• 4.Sausville, E.A., Arbuck, S.G., Messmann, R., Headlee, D., Bauer, K.S., Lush, R.M., Murgo, A., Figg, W.D., Lahusen, T., Jaken, S., Jing, X., Roberge, M., Fuse, E., Kuwabara, T. & Senderowicz, A.M. (2001) Phase I trial of 72-hour continuous infusion UCN-01 in patients with refractory neoplasm. J. Clin. Oncol. 19, 2319-2333.
• 5.Senderowicz, A.M. & Sausville, E.A. (2000) Preclinical and clinical development of cyclin- dependent kinase modulators. J. Natl. Cancer Inst. 92, 376-387.
• 6.Taylor, W.R. & Stark, G.R. (2001) Regulation of the G2/M transition by p53. Oncogene20, 1803-1815.
• 7.Lee, J.M. & Bernstein, A. (1993) p53 mutations increase resistance to ionizing radiation. Proc. Natl. Acad. Sci. U.S.A. 90, 5742-5746.
• 8.Fan, S., el-Deiry, W.S., Bae, I., Freeman, J., Jondle, D., Bhatia, K., Fornace, A.J., Jr., Magrath, I., Kohn, K.W. & O'Connor, P.M. (1994) p53 gene mutations are associated with decreased sensitivity of human lymphoma cells to DNA damaging agents. Cancer Res. 54, 5824-5830.
• 9.Fan, S., Smith, M.L., Rivet, D.J. 2nd, Duba, D., Zhan, Q., Kohn, K.W., Fornace, A.J., Jr. & O'Connor, P.M. (1995) Disruption of p53 function sensitizes breast cancer MCF-7 cells to cisplatin and pentoxifylline. Cancer Res. 55, 1649-1654.
• 10.Smith, M L., Kontny, H.U., Zhan, Q., Sreenath, A., O'Connor, P.M. & Fornace, A.J., Jr. (1996) Antisense GADD45 expression results in decreased DNA repair and sensitizes cells to u.v.-irradiation or cisplatin. Oncogene 13, 2255-2263.
• 11.Cimoli, G., Valenti, M., Ottoboni, C. & Parodi, S. (2001) HPV16-E6 enhances mitoxantrone sensitivity in a human ovarian cancer line: An isolated instance or a trend? Int. J. Oncol. 18, 759-765.
• 12.Bunz, F., Dutriaux, A., Lengauer, C., Waldman, T., Zhou, S., Brown, J.P., Sedivy, J.M., Kinzler, K.W. & Vogelstein, B. (1998) Requirement for p53 and p21 to sustain G2 arrest after DNA damage. Science 282, 1497-1501.
• 13. Skladanowski, A. & Larsen, A.K. (1997) Expression of wild-type p53 increases etoposide cytotoxicity in Ml myeloid leukemia cells by facilitated G2 to M transition: Implications for gene therapy. Cancer Res. 57, 818-823. 14. Skladanowski, A. & Konopa, J. (1993) Adriamycin and daunomycin induce programmed cell death (apoptosis) in tumour cells. Biochem. Pharmacol. 46, 375-382.
• 15.el-Deiry, W.S., Harper, J.W., O'Connor, P.M., Velculescu, V.E., Canman, C.E., Jackman, J., Pietenpol, J.A., Burrell, M., Hill, D.E., Wang, Y., Wiman, K.G., Mercer, W.E., Kastan, M.B., Kohn, K.W., Elledge, S.J., Kinzler, K.W. & Vogelstein, B. (1994) WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res. 54, 1169-1174.
• 16.Guillouf, C., Rosselli, F., Krishnaraju, K., Moustacchi, E., Hoffman, B. & Liebermann, D.A. (1995) p53 involvement in control of G2 exit of the cell cycle: Role in DNA damage-induced apoptosis. Oncogene 10, 2263-2270.
• 17.Wang, Q., Fan, S., Eastman, A., Worland, P.J., Sausville, E.A. & O'Connor, P.M. (1996) UCN-01: A potent abrogator of G2 checkpoint function in cancer cells with disrupted p53. J. Natl. Cancer Inst. 88, 956-965.
• 18.Borbe, R., Rieger, J. & Weller, M. (1999) Failure of taxol-based combination chemotherapy for malignant glioma cannot be overcome by G2/M checkpoint abrogators or altering the p53 status. Cancer Chemother. Pharmacol. 44, 217-227.
• 19.Sugiyama, K., Shimizu, M., Akiyama, T., Tamaoki, T., Yamaguchi, K., Takahashi, R., Eastman, A. & Akinaga, S. (2000) UCN-01 selectively enhances mitomycin C cytotoxicity in p-53 defective cells which is mediated through S and/or G(2) checkpoint abrogation. Int. J. Cancer 85, 703-709.
• 20.Shao, R.G., Cao, C.X., Shimizu, T., O'Connor, P.M., Kohn, K.W. & Pommier, Y. (1997) Abrogation of an S-phase checkpoint and potentiation of camptothecin cytotoxicity by 7-hydroxystaurosporine (UCN-01) in human cancer cell lines, possibly influenced by p53 function. Cancer Res. 57, 4029-4036.
• 21.Jones, C.B., Clerments, M.K., Wasi, S. & Daoud, S.S. (2000) Enhancement of camptothecin-induced cytotoxicity with UCN-01 in breast cancer cells: Abrogation of S/G(2) arrest. Cancer Chemother. Pharmacol. 45, 252-258.
• 22.Jones, C.B., Clements, M.K., Redkar, A. & Daoud, S.S. (2000) UCN-01 and camptothecin induce DNA double-strand breaks in p53 mutant tumor cells, but not in normal or p53 negative epithelial cells. Int. J. Oncol. 17, 1043-4051.
• 23.Smith, ML., Chen, I.T., Zhan, Q., O'Connor, P.M. & Fornace, A.J., Jr. (1995) Involvement of the p53 tumor suppressor in repair of u.v.-type DNA damage. Oncogene 10, 1053-1059.
• 24.Kartalou, M. & Essigmann, J.M. (2001) Mechanisms of resistance to cisplatin. Mutat. Res. 478, 23-43.
• 25.Li, T.-K. & Liu, L.F. (2001) Tumor cell death induced by topoisomerase-targeting drugs. Annu. Rev. Pharmacol. Toxicol. 41, 53-77.
• 26. Sonoda, E., Takata, M., Yamashita, Y.M., Morrison, C. & Takeda, S. (2001) Homologous DNA recombination in vertebrate cells. Proc. Natl. Acad. Sci. U.S.A. 98, 8388-8394.
• 27. Karran, P. (2000) DNA double strand breaks repair in mammalian cells. Curr. Opin. Genet. Dev. 10, 144-150.
• 28.Goldwasser, F., Shimizu, T., Jackman, J., Hoki, Y., O'Connor, P.M., Kohn, K.W. & Pommier, Y. (1996) Correlations between S and G2 arrest and the cytotoxicity of camptothecin in human colon carcinoma cells. Cancer Res. 56, 4430-4437.
• 29.Bates, S. & Vousden, K.H. (1996) p53 in signaling checkpoint arrest or apoptosis. Curr. Opin. Genet. Dev. 6, 1-7.