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目的探讨RNAi靶向沉默对人卵巢癌OVCAR3细胞CD105和Ki67基因的表达。方法采用脂质体介导的基因法将不同浓度的CD105-siRNA、Ki67-siRNA转染至体外培养的人卵巢癌OVCAR3细胞中,检测干扰前后人卵巢癌细胞中CD105、Ki67基因表达的变化以确定沉默效果。构建CD105-siRNA、Ki67-siRNA及各自的阴性对照序列并高效地转染人卵巢癌OVCAR3细胞,采用Western blotting和Real-time PCR从蛋白和基因水平检测CD105和Ki67的表达变化。结果转染CD105-siRNA或Ki67-siRNA终浓度为50nmol/L和100nmol/L的人卵巢癌细胞中CD105mRNA或Ki67mRNA的相对表达水平及CD105和Ki67的表达均明显低于空白对照组和阴性对照组(NC1),差异具有统计学意义(P<0.001)。结论 CD105-siRNA和Ki67-siRNA能特异性抑制人卵巢癌OVCAR3细胞中的CD105和Ki67基因表达,下调mRNA和蛋白的表达水平。
Objective To investigate the expression of CD105 and Ki67 in human ovarian cancer OVCAR3 cells by RNAi targeted silencing. Methods Different concentrations of CD105-siRNA and Ki67-siRNA were transfected into human ovarian cancer OVCAR3 cells by liposome-mediated gene method to detect the expression of CD105 and Ki67 in human ovarian cancer cells before and after interference Determine the effect of silence. CD105-siRNA, Ki67-siRNA and their respective negative control sequences were constructed and efficiently transfected into ovarian cancer OVCAR3 cells. The expressions of CD105 and Ki67 were detected by Western blotting and Real-time PCR at the protein and gene level. Results The relative expression levels of CD105mRNA or Ki67mRNA and the expressions of CD105 and Ki67 in human ovarian cancer cells transfected with CD105-siRNA or Ki67-siRNA at final concentration of 50nmol / L and 100nmol / L were significantly lower than those of blank control group and negative control group (NC1), the difference was statistically significant (P <0.001). Conclusion CD105-siRNA and Ki67-siRNA can specifically inhibit the expression of CD105 and Ki67 in ovarian cancer OVCAR3 cells and down-regulate mRNA and protein expression levels.