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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。
AIM: To investigate the effect of small interfering RNA (siRNA) on the proliferation of cervical cancer HeLa cells by inhibiting the expression of Ca2 + -activated K + channels (IKCa1). Methods: The siRNA expression vector containing IKCa1 gene was constructed and transfected into HeLa cells by lipofectamine. The mRNA and protein expression of IKCa1 in HeLa cells was detected by RT-PCR and Western Blot. The proliferation of HeLa cells was detected by flow cytometry HeLa cell cycle changes. Results: The eukaryotic expression vector targeting IKCa1 gene was successfully constructed. The expression of IKCa1 mRNA and protein in HeLa cells was significantly decreased (P <0.05) and inhibited HeLa cell proliferation (P <0.05) Cell cycle stays in the G0 ~ G1 phase, S phase cells decreased significantly. Conclusion: The siRNA interference technique can effectively inhibit the expression of IKCa1 gene and inhibit the proliferation activity of cervical cancer HeLa cells in vitro.