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The Effect of Anti-HPV16 E6-Ribozyme on Sensitivity of Cervical Carcinoma Cell Line to Cisplatin
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Abstract:
To investigate the effect of anti-HPV16 E6-ribozyme on sensitivity of cervical carcinoma cells line to cisplatin. Methods: With the method of lipofectin transfection, the anti-HPV16E6-ribozyme and the eucaryotic expressing plasmids per se were transfected into CaSKi cells, which were named CaSKi-R, CaSKi-P respectively. The expression of ribozyme in transfected cells was observed by RNA dot blot. The amounts of E6 mRNA in the three kinds of cells were detected by Northern blot. The growth curve and colony formation test of three cell types were detected. Their sensitivity to cisplatin were examined by MTT assay. The apoptosis rates of each cell and expressions of P53, Bcl-2 and Bax was determined by PI/Annexin V stained methods and flow cytometry analysis.Results: Anti-HPV16E6-ribozyme can be expressed stably in transfected CaSKi cells. Northern blot showed that E6 mRNA was less in CaSKi-R than in CaSKi. The growth rate and cloning efficiency of CaSKi-R cells were lower than that of CaSKi and CaSKi-P cells. The sensitivity and apoptotic rates of CaSKi-R cells to cisplatin were higher than that of CaSKi and CaSKi-P cells( P <0.01).The expression of P53, Bax protein was upregulated and the expression of Bcl-2 protein downregulated compared to that of CaSKi and CaSKi-P cells( P <0.01).Conclusion: Anti-HPVE6-ribozyme inhibited CaSKi-R cell growth, cloning efficiency and increased the sensitivity to cisplatin.
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