RNA干扰对SKOV3卵巢癌细胞株Skp2表达的抑制作用

• 论著 •

RNA干扰对SKOV3卵巢癌细胞株Skp2表达的抑制作用

齐凤平¹,潘国强²

1. 迁安市妇幼保健院
2. 迁安市人民医院

收稿日期:2009-10-14 修回日期:2010-04-23 出版日期:2010-09-15 发布日期:2010-09-15
通讯作者: 潘国强

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

Received:2009-10-14 Revised:2010-04-23 Published:2010-09-15 Online:2010-09-15

齐凤平1 ,潘国强2 . RNA干扰对SKOV3卵巢癌细胞株Skp2表达的抑制作用[J]. .

摘要/Abstract

摘要： 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，揭示Skp2与细胞增殖的关系。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、阴性对照组及阳性对照组。采用Realtime PCR和Western Blotting技术检测各组Skp2 mRNA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，采用方差分析进行组间均数比较，以p<0.05表示差异有统计学意义。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Skp2 mRNA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），结论：将人Skp2-siRNA转染入SKOV3卵巢癌细胞株后，能够抑制SKOV3卵巢癌细胞株Skp2表达，从而抑制卵巢癌细胞的增殖。

关键词: S期激酶相关蛋白2, RNA干涉细胞, 周期调控

Abstract: Objective To investigate suppression effect of RNA interference (RNAi) on expression of Skp2 of SKOV3 ovarian carcinoma cell line, The research is expected to reveal the relationship between Skp2 and cell cycle regulation and establish the theory basis for target gene therapy. Methods The Skp2-siRNA was designed and synthesized by Shanghai Gene Pharma Co., Ltd，and then transfected to SKOV3 ovarian carcinoma cell line. The cell line were divided into six groups, including the blank control group, the siRNA transfection group 1, the siRNA transfection group 2, the mock transfection group, the negative control group and the positive 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 line was observed with CCK-8 assay, results were analyzed statistically by ANOVA. Results After transfection with Skp2-siRNA, the expression level of Skp2 mRNA and protein in the siRNA transfection group 1 and group 2 were down-regulated, the growth and proliferation of SKOV3 Ovarian Carcinoma Cell line in the siRNA transfection group 1 and the siRNA transfection group 2 were inhibited, There were difference in the siRNA transfection group 1 and group 2 from in the blank control group (p<0.05), but no statistics difference between in the siRNA transfection group 1 and in the siRNA transfection group 2 (p>0.05). Conclusions 　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 gene silencing of oncogene.

Key words: Skp2, RNA interfering, cell cycle regulation