Cancer Research and Treatment

The Korean Cancer Association (대한암학회)
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Cell Cycle Regulatory Protein Expression Profiles by Adenovirus p53 Infection in Human Papilloma Virus-associated Cervical Cancer Cells

Lee, Yong-Seok;Bae, Su-Mi;Kwak, Sun-Young;Park, Dong-Chun;Kim, Yong-Wook;Hur, Soo-Young;Park, Eun-Kyung;Han, Byoung-Don;Lee, Young-Joo;Kim, Chong-Kook;Kim, Do-Kang;Ahn, Woong-Shick

  • Published : 20060000
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Abstract

Purpose: The tumor suppressor gene, p53, has been established as an essential component for the suppression of tumor cell growth. In this study, we investigated the time-course anticancer effects of adenoviral p53 (Adp53) infection on human ovarian cancer cells to provide insight into the molecular-level understanding of the growth suppression mechanisms involved in Adp53-mediated apoptosis and cell cycle arrest. Materials and Methods: Three human cervical cancer cell lines (SiHa, CaSki, HeLa and HT3) were used. The effect of Adp53 infection was studied via cell count assay, cell cycle analysis, FACS, Western blot and macroarray assay. Results: Adp53 exerts a significant role in suppressing cervical cancer cell growth. Adp53 also showed growth inhibitory effects in each cell line, and it induced apoptosis and cell cycle arrest. Adp53 differentially regulated the expression of genes and proteins, and the gene expression profiles in the SiHa cells revealed that the p21, p53 and mdm2 expressions were significantly up-regulated at 24 and 48 hr. Western blot shows that the p21 and p53 expressionlevels were significantly increased after Adp53 infection. In addition, in all cell lines, both the CDK4 and PCNA protein expression levels were decreased 48 h after Adp53 infection. Cell cycle arrest at the G1 phase was induced only in the SiHa and HeLa cells, suggesting that exogenous infection of Adp53 in cancer cells was significantly different from the other HPV- associated cervical cancer cells. Conclusion: Adp53 can inhibit cervical cancer cell growth through induction of apoptosis and cell cycle arrest, as well as through the regulation of the cell cycle-related proteins. The Adp53-mediated apoptosis can be employed as an advanced strategy for developing preferential tumor cell-specific delivery. (Cancer Res Treat. 2006;38:168-177)

Keywords

References

  1. Vozelstein B, Kinzler KW. p53 function and dysfunction. Cell. 1992;70:523-6 https://doi.org/10.1016/0092-8674(92)90421-8
  2. Buckbinder L. Talbott R. Seizinzer BR. Klev N. Gene regulation by temperature-sensitive p53 mutants: identification of p53 response genes. Proc Natl Acad Sci USA. 1994;91: 10640-4
  3. Hamada K, Alemanv R. Zhang WW, Hittelman WN, Lotan R. Roth JA. et al. Adenovirus-mediated transfer of a wild-type p53 gene and induction of apoptosis in cervical cancer. Cancer Res. 1996;56:3047-54
  4. Huang TG, Ip SM, Yeung WS, Ncan HY. Changes in p21WAF1, pRb, Mdm-Z, Bax and Bcl-2 expression in cervical cancer cell lines transfected with a p53 expressing adenovirus. Eur J Cancer. 2000;36:249-56 https://doi.org/10.1016/S0959-8049(99)00247-6
  5. Polyak K, Waldman T, He TC, Kinzler KW, Voaelstein B. Genetic determinants of p53-induced apoptosis and growth arrest. Genes Dev. 1996;10:1945-52 https://doi.org/10.1101/gad.10.15.1945
  6. Shillitoe EJ, Kamath P, Chen Z. Panillomaviruses as targets for cancer gene therapy. Cancer Gene Ther. 1994;1:193-204
  7. Kaur P, Mcfrouaall JK, Cone R. Immortalization of primary human epithelial cells by cloned cervical carcinoma DNA containing human papillomavirus type 16 E6{E7 open reading frames. J Gen Virol. 1989;70:1261-6 https://doi.org/10.1099/0022-1317-70-5-1261
  8. Mclvlurrav HR Nguyen D, Westbrook TF, McAnce DJ. Biology of human papillornaviruses. Int J Exp Pathol. 2001;82: 15-33 https://doi.org/10.1046/j.1365-2613.2001.00177.x
  9. Lee SJ, Namkoong SE, Lee WC, Sui JW, Jee SH, You YK, et al. Polvmornhisms of p53, p21 and IRF-l and cervical cancer susceptibility in Korean women. Cancer Res Treat. 2002; 34:357-64 https://doi.org/10.4143/crt.2002.34.5.357
  10. Hamada K, Zhang WW, Alemanv R. Wolf J, Roth JA, Mitchell MF. Growth inhibition of human cervical cancer cells with the recombinant adenovirus p53 in vitro. Gynecol Oncol. 1996;60:373-9 https://doi.org/10.1006/gyno.1996.0057
  11. Graham FL, Prevec L. Methods for construction of adenovirus vectors. Mol Biotechnol. 1995;3:207-20 https://doi.org/10.1007/BF02789331
  12. Xu HI, Zhou Y, Seizne J, Perna GS, Mixon M, Zhang C, et al. Enhanced tumor suppressor gene therapy via replicationdeficient adenovirus vectors expressing an N-terminal truncated retinoblastoma protein. Cancer Res. 1996;56:2245-9
  13. Ahn WS, Han YI, Bae SM, Kim TH, Rho MS, Lee JM, et al. Differential suppression of human cervical cancer cell growth by adenovirus delivery of p53 in vitro: arrest phase of cell cycle is dependent on cell line. Jpn J Cancer Res. 2002; 93:1012-9 https://doi.org/10.1111/j.1349-7006.2002.tb02478.x
  14. Ahn WS, Bae SM, Lee JM, Narnkoonz SE, Yoo JY, Seo YS, et al. Anti-cancer effect of adenovirus p53 on human cervical cancer cell growth in vitro and in vivo. Int J Gynecol Cancer. 2004;14:322-32 https://doi.org/10.1111/j.1048-891x.2004.014217.x
  15. Ahn WS, Bae SM, Lee KH, Lee JM, Namkoong SE, Chun HJ, et al. Recombinant adenovirus-p53 gene transfer and cellspecifie growth suppression of human cervical cancer cells in vitro and in vivo. Gvnecol Oncol. 2004;92:611-21 https://doi.org/10.1016/j.ygyno.2003.10.033
  16. Scheffner M, Munger K, Byrne JC, Howley PM. The state of the p53 and retinoblastoma genes in human cervical carcinoma cell lines. Proc Natl Acad Sci. 1991;88:5523-7 https://doi.org/10.1073/pnas.88.13.5523
  17. Harbour JW, Luo RX, Dei Santi A, Postigo AA, Dean DC.Cdk phosphorylation triggers sequential intramolecular interactions that progressively block Rb functions as cells move through GI. Cell. 1999;98:859-69 https://doi.org/10.1016/S0092-8674(00)81519-6
  18. Rogoff HA, Pickering MT, Frame FM, Debatis ME, Sanchez Y, Jones S, et al. Apoptosis associated with deregulated E2F activity is dependent on E2Fl and Atrn/Nbsl/Chk2. Mol Cell BioI. 2004;24:2968-77 https://doi.org/10.1128/MCB.24.7.2968-2977.2004
  19. Hall PA, Levison DA, Woods AL, Yu CC, Kellock DB, Watkins JA, et al. Proliferating cell nuclear antigen immunolocalization in paraffin sections: an index of cell proliferation with evidence of deregulated expression in some neoplasms. J Pathol. 1990;162:285-94 https://doi.org/10.1002/path.1711620403
  20. Polager S, Ginsberg D. E2F mediates sustained G2 arrest and down-regulation of Stathmin and AIM-l expression in response to genotoxic stress. J Bioi Chem. 2003;278:1443-9 https://doi.org/10.1074/jbc.M210327200
  21. Lizard G, Chignol MC, Chardonnet Y, Schmitt D. Differences of reactivity to interferon gamma in HeLa and CaSki cells: a combined immunocytochemical and t1ow-cytometric study. J Cancer Res Clin Oncol. 1996;122:223-30 https://doi.org/10.1007/BF01209650
  22. Chen X, Ko LJ, Javaraman L, Prives C. p53 levels, functional domains, DNA damage determine the extent of the apoptotic response of tumor cells. Gene Dev. 1996;10:2438-51 https://doi.org/10.1101/gad.10.19.2438
  23. Shao J, Fujiwara T, Kadowaki Y, Fukazawa T, Waku T, Itoshima T, et al. Overexpression of the wild-type p53 gene inhibits NF-kappaB activity and svneraizes with aspirin to induce apoptosis in human colon cancer cells. Oncogene. 2000; 19:726-36 https://doi.org/10.1038/sj.onc.1203383
  24. Robles AI, Bemmels NA, Foraker AB, Harris CC. APAF-l is a transcriptional target of p53 in DNA damage-induced apoptosis. Cancer Res. 2001;61:6660-4