Influence of survivin short hairpin RNA on cell proliferation and apoptosis in cervical cancer cell line HeLa

Objective:To investigate the influence of survivin short hairpin RNA(shRNA) on cell proliferation and apoptosis in cervical cancer cell line HeLa.Methods:Having transfected the recombined eukaryotic expression plamid pSilencer 2.1-s2 which concerned human survivin gene shRNA into human cervical cancer cell line HeLa by using LipofectAMINE2000,then selected the positive clones with G418,detected the expression of survivin mRNA and its protein by semi-quantitative RT-PCR and Western blot respectively,protracted cell growth curve by methyl thiazolyl tetrazolium(MTT) assay,examined the changes of cell cycle distribution by flow cytometry,observed cell apoptosis by flow cytometry and Hoechst staining.Results:Obtained the positive clones by screening with G418 for 34 days,compared with the cells which having been transfected with negative control plasmid(HeLa-NC),pure plasmid(HeLa-U6 neo)and those not having been transfected(HeLa),the expression level of survivin gene mRNA and protein declined evidently in the cells which having been transfected with pSilencer2.1-s2 plasmid(HeLa-s2),the expression inhibitory rate were 62.8% and 60.1%;the cell proliferation was inhibited,and the highest inhibitory rate was(57.8±2.1)%(P0.05);the changes of cell cycle distribution were significant,many cells were blocked in G_0/G_1 phase(72.7±3.1)%(P0.05),G_2/M phase(5.1±2.9)% reduced sharply(P0.05);the apoptotic rate was(29.2±1.4)%,rising up obviously(P0.05).Conclusion:Survivin gene shRNA can reduce the expression of survivin in human cervical cancer cell line HeLa,inhibit cell proliferation and induce cell apoptosis.