Down Regulation of Survivin Gene and Up Regulation of p53 Gene expression by siRNA Induces Apoptosis in human Hepatocellular Carcinoma cell Line HepG2
- 1 Research Centre of Biotechnology, China Pharmaceutical University, NanJing, 210009, China
- 2 Research Centre of Biotechnology, China Pharmaceutical University, NanJing, 210009, China
- 3 Research Centre of Biotechnology, China Pharmaceutical University, NanJing
- 4 Research Centre of Biotechnology, China Pharmaceutical University, NanJing, 210009, China
Abstract
Survivin gene may be a good target for cancer gene therapy because it is over expressed in a variety of human tumors including human hepatocellular carcinoma but not in differen- tiated adult tissues. To explore the effects of the siRNA of survivin gene inducing apoptosis in human hepatocellular cancer cells, three siRNAs cpusiRNA1, cpusiRNA2 and cpusiRNA3 were designed and transferred into human hepatocellular carcinoma cell line HepG2 (HepG2) by lipofection. MTT test showed that the growth of HepG2 decreased when it was transfected with 25nM, 50nM, 100nM, 150nM, 200nM, 400nM siRNA respectively after 48 hours. And the change of mRNA and protein of survivin gene and p53 gene had been detected by RT-PCR and Western blot. Cells presented an increase in apoptosis index was assayed by flow cytometry. Small interfering RNA can exert a knockdown of survivin gene expression and up regulation of p53 gene to induce apoptosis and to inhibit the growth of HepG2.
- H. Caldas, L. E. Honsey and R. A. Altura. (2005) Survivin a novel Survivin splice variant expressed in malignancies. Mol Cancer, 4(1): 11-23.
- W. Salz, D. Eisenberg and J. Plescia. (2005) A survivin gene signature predicts aggressive tumor behavior. Cancer Res, 65: 3531-3534.
- T. Liu and D. Grossman. (2004) Rapid induction of mitochondria events and caspase-independent apoptosis in Survivin-targeted melanoma cells. Oncogene, 23 (1): 39-48.
- M. Stevenson. (2004) Therapeutic potential of RNA interference. N Engl J Med, 351 (17): 1772-1777.
- D. C. Altieri; (2003) Survivin versatile modulation of cell division and apoptosis in cancer. Oncogene, 22: 8581–8589.
- X. Ling, R. J. Bernacki, M. G. Brattain and F. Li. (2004) Induction of survivin expression by taxol (paclitaxel) is an early event, which is independent of taxol-mediated G2/M arrest. J Biol Chem; 279: (5)196-203.
- C. D. Novina and P. A. Sharp. (2004) The RNAi revolution. Na-ture, 430(6996): 161-164.
- P. D. Zamore. (2002) Ancient pathways p rogrammed by small RNAs. Science, 296 (5571): 1265-1269.
- G. J. Hannon. (2002) RNA interference. Nature, 418: 2442-2511.
- M. Pennati, M. Binda, M. De Cesare, G. Pratesi, M. Folini, L. Citti, M. G. Daidone, F. Zunino and N. Zaffaroni. (2004) Ri-bozyme-mediated down-regulation of survivin expression sensi-tizes human melanoma cells to topotecan in vitro and in vivo. Carcinogenesis; 25: 1129-1136.
- M. Izquierdo. (2005) Short interfering RNAs as a tool for cancer gene therapy. Cancer Gene Ther, 12: 217-227.
- M. Kappler, M. Bache, F. Bartel, M. Kotzsch, M. Panian, P. Wurl, K. Blumke, H. Schmidt, A. Meye and H. Taubert. (2004) Knock-down of survivin expression by small interfering RNA reduces the clonogenic survival of human sarcoma cell lines independ-ently of p53. Cancer Gene Ther, 11: 186-193.
- M .Kappler, H. Taubert, F. Bartel, K. Blumke, M. Panian, H. Schmidt, J. Dunst and M. Bache. (2005) Radiosensitization, after a combinedtreatment of survivin siRNA and irradiation, is correlated with the activation of caspases 3 and 7 in a wtp53 sarcoma cell line, but not in a mt-p53 sarcoma cell line. Oncol Rep, 13: 167-172.
- M. Chawla-Sarkar, S. I. Bae, F. J. Reu, B. S. Jacobs, D. J. Lindner and E. C. Borden. (2004) Downregulation of Bcl-2, FLIP or IAPs (XIAP and survivin) by siRNAs sensitizes resistant melanoma cells to Apo2L/TRAIL-induced apoptosis. Cell Death Differ, 11: 915-923.