siRNA抑制IKCa1基因表达对宫颈癌HeLa细胞增殖的影响

目的:研究siRNA(small interfering RNA)抑制钙激活性中电导钾离子通道(intermediate-conductance Ca2+-activated K+channels,IKCa1)表达对宫颈癌HeLa细胞增殖的影响。方法:构建含有IKCa1基因的siRNA表达载体,脂质体法转染HeLa细胞,通过RT-PCR和Western Blot检测HeLa细胞IKCa1 mRNA和蛋白表达变化;WST-1检测细胞增殖能力;流式细胞仪分析HeLa细胞周期变化。结果:成功构建针对IKCa1基因的真核表达载体,所获表达载体转染HeLa细胞可使IKCa1mRNA和蛋白表达水平显著降低(P<0.05),并能有效抑制HeLa细胞增殖(P<0.05),使细胞周期停留在G0～G1期,S期细胞明显减少。结论:应用siRNA干扰技术能有效抑制IKCa1基因的表达,同时也可有效抑制宫颈癌HeLa细胞体外增殖活性。

Effect of IKCa1 gene expression inhibited by siRNA on the proliferation of cervical cancer HeLa cells

Objective:To study the effect of intermediate-conductance Ca2+-activated K+ channels(IKCa1) gene expression inhibited by small interfering RNA(siRNA) on the proliferation of cervical cancer HeLa cells.Methods:siRNA expression vector containing IKCa1 gene was constructed,HeLa cells were transfected by liposome method,RT-PCR and Western Blot were used to detect the expressions and changes of IKCa1 mRNA and protein in HeLa cells;the proliferative ability of HeLa cells was detected by WST-1;flow cytometry was used to analyze the cycle change of HeLa cells.Results:The eukaryotic expression vector targeting to IKCa1 gene was constructed successfully.The expression levels of IKCa1 mRNA and protein decreased significantly after transfecting the expression vector into HeLa cells(P<0.05),and the proliferation of HeLa cells was inhibited effectively(P<0.05).The cell cycle was stopped at G0-G1 phase,the number of HeLa cells in S phase decreased significantly.Conclusion:siRNA interfering technique can inhibit the expression of IKCa1 gene and proliferative activity of cervical cancer HeLa cells in vitro effectively.