靶向survivin基因siRNA对喉癌细胞增殖、凋亡的影响

Investigation of siRNA targeted to survivin in uppressing proliferation and inducing apoptosis in laryngeal carcinoma

目的观察靶向survivin基因的siRNA对喉癌细胞Hep-2增殖和凋亡的影响,为喉癌的基因治疗提供有效靶点及技术。方法使用阳离子脂质体LipofectamineTM2000将靶向survivin基因的siRNA转染至喉癌细胞株Hep-2;MTT[3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐]法检测对喉癌细胞Hep-2增殖的影响;RT-PCR检测喉癌细胞Hep-2survivin基因mRNA表达的变化;Western Blot检测survivin基因蛋白表达;流式细胞技术检测喉癌细胞Hep-2凋亡情况。结果各浓度survivin-siRNA转染组细胞增殖抑制率和凋亡率均明显高于对照组(<0.05),survivinmRNA和蛋白表达有不同程度减弱,并且有明显的浓度依赖性,随着浓度的上升表达率下降。结论利用RNA干扰技术沉默survivin基因表达可以显著抑制喉癌细胞Hep-2细胞增殖,并在一定程度上诱导其自发凋亡,不同浓度survivin-siRNA转染能下调survivinmRNA和蛋白表达,靶向survivin基因的RNA干扰技术在喉癌的基因治疗中具有一定价值。

Objective To investigate the effect of blocking the survivin expression with RNAinterference（RNAi） techniques on suppressing proliferation and inducing apoptosis of laryngeal carcinoma Hep-2 cells,and supply an effective target and technique for laryngeal carcinoma gene therapy.Methods A siRNA directed against survivin was transfected into laryngeal carcinoma Hep-2 cells with lipofectamineTM2000.The effect of suppressing proliferation of Hep-2 cell was detected by MTT assay.The changes of survivin expression were detected by semiquantitive RT-PCR.The protein of survivin was evaluated by means of Western blot.The effect of inducing Hep-2 cell apoptosis was detected by flow cytometry.Results The inhibition ratio of proliferation and the apoptotic ratio were significantly higher than those of control grou（0.05）,Expressions of survivin gene in mRNA and protein levels were suppressed by survivin-siRNA in a dose-dependent manner.Conclusion Blocking the survivin expression with RNAi technology can significantly suppress proliferation of Hep-2 cells and induce apoptosis to a certain degree.Expressions of survivin mRNA and protein in different dose siRNA groups are decreased in different degrees.RNAi targeted to survivin has a potential value in gene therapy of laryngeal carcinoma.