联合应用以Hif-1α和Survivin基因为靶标的siRNA对鼻咽癌CNE-2细胞增殖和周期的影响

Combined application the siRNA target for Hif-1α and Survivin gene to human NPC CNE-2 cell and its effects on the proliferation and cycle of this cell

目的:研究联合转染以乏氧诱导因子-1α(Hif-1α)和Survivin基因为靶标的siRNA后,鼻咽癌CNE-2细胞增殖和周期的改变。方法:以Hif-1α和Survivin基因为靶标的siRNA共同转染鼻咽癌CNE-2细胞,以单靶点siRNA转染组和未处理组为对照。MTT检测转染后细胞的增殖活力;流式细胞仪检测转染后细胞的周期变化,RT-PCR与Western Blot技术检测转染后各组细胞相应靶基因mRNA及相关蛋白表达水平的变化。结果:鼻咽癌CNE-2细胞分别转染以Hif-1α和Survivin基因为靶标的siRNA后,细胞的增殖能力明显下降,相应靶基因的mRNA及蛋白的表达明显减低,细胞周期出现G0/G1期阻滞;联合转染两种siRNA后细胞的各项检测结果均明显强于对照组,与各组相比差异均有统计学意义(P<0.01)。结论:联合以Hif-1α和Survivin基因为靶标的siRNA干扰组,可以较单基因干扰组更有效干扰鼻咽癌CNE-2细胞内靶基因mRNA和蛋白的表达,抑制细胞的增殖。

Objective：To study the combined transfection of the siRNA target for Hif-1α and Survivin gene to human NPC CNE-2 cell and its effects on the proliferation and cycle of this cell. Method：Combined transfection of the siRNA target for Hif-1α and Survivin gene to human NPC CNE-2 cell, these plasmids were respectively trans- fected into the same cells. Cell proliferation was detected with MTT assay. The inhibitory effects on target genes were evaluated with RT-PCR and Western Blot at the levels of mRNA and protein, respectively. Result.. MTT showed that CNE-2 cell proliferation in multi-gene plasmid group was more significantly inhibited than a single gene. The expression of mRNA and protein of two different genes were both decreased in HS group, and the in- terference effect of multiple genes was better than the single-gene（P〈0. 01）. Conclusion： HS group could restrain cell proliferation and interference the mRNA and protein expression in nasopharyngeal carcinoma CNE-2 cell, which was better than the other groups.