RNA干扰对SKOV3卵巢癌细胞株Skp2表达的抑制作用

Suppression of RNA Interference on Expression of Skp2 of SKOV3 Ovarian Carcinoma Cell Line

目的:探讨RNA干扰(RNAi)技术对SKOV3卵巢癌细胞株Skp2表达的抑制作用。方法:设计合成针对Skp2的小干涉RNA(siRNA)并进行转染。实验分为空白对照组、转染组1、转染组2、转染对照组及阴性对照组。采用Real time PCR和Western blotting技术检测各组Skp2mRNA和蛋白水平的变化,通过细胞计数试剂盒-8(CCK-8)法检测各组卵巢癌细胞增殖情况。结果:应用Skp2-siRNA干扰SKOV3卵巢癌细胞株后,转染组1、转染组2的Skp2mRNA和蛋白表达水平明显降低,细胞增殖明显受限,与空白对照组相比较,差异有统计学意义(P<0.05),转染组1与转染组2之间差异无统计学意义(P>0.05)。转染组1与转染组2基因沉默效率分别为75.31%和76.86%,蛋白抑制率分别为62.10%和63.11%,细胞增殖抑制率分别为52.75%和53.06%。结论:RNAi技术能够抑制SKOV3卵巢癌细胞株Skp2的表达,从而抑制卵巢癌细胞的增殖。

Objective： To investigate suppression of RNA interference （RNAi） on expression of Skp2 of SKOV3 ovarian carcinoma cell line. Methods： The Skp2-siRNA was designed and synthesized, then transfected to SKOV3 ovarian carcinoma cell lines. The cell lines were divided into five groups, including the blank control group, the siRNA transfection group 1, the siRNA transfection group 2, the mock transfection group and the negative control group. The expression level of Skp2 mRNA and protein were detected by realtime-PCR and Western blotting, respectively. The growth and proliferation of SKOV3 ovarian carcinoma cell lines were observed with CCK-8 assay. Results： After transfection with Skp2-siRNA, the expression level of Skp2 mRNA and protein were down-regulated, the growth and proliferation of SKOV3 ovarian carcinoma cell line were inhibited in the siRNA transfection group 1 and the siRNA transfection group 2. There were significant differences between the siRNA transfection group 1 and group 2 and the blank control group（P 0.05）,but no significant difference between the siRNA transfection group 1 and siRNA transfection group 2 （P 0.05）. The silencing efficiencies were 75.31% and 76.86% in siRNA transfection group 1 and group 2 respectively. The protein suppression rates were 62.10% and 63.11% respectively. The inhibition ratios were 52.75% and 53.06% respectively by CCK-8 assay. Conclusion： The down-regulation of Skp2 expression of SKOV3 ovarian carcinoma cell line by Skp2-siRNA can lead to the suppression of proliferation. The small interfering RNAs technique can inhibit the proliferation of carcinoma cell by oncogene silencing.