RNAi技术抑制HepG2细胞中DcR3的表达

目的探讨特异性DcR3-siRNA对人肝细胞癌细胞系HepG2的DcR3基因表达的影响。方法设计并合成带FAM荧光标记的针对DcR3的siRNA4条和非特异性siRNA1条,用脂质体转染HepG2细胞,应用荧光显微镜和流式细胞仪判断转染效果;应用半定量RT-PCR和免疫细胞化学检测特异性DcR3-siRNA的对HepG2细胞中DcR3表达的抑制作用。结果各特异性DcR3-siRNA均能抑制DcR3 mRNA表达(P<0.05),其中以siRNA4的作用更明显,干扰作用达62.9%;将siRNA4转染HepG2细胞,免疫细胞化学结果显示,siRNA4能明显抑制HepG2细胞中DcR3蛋白的表达(P<0.01)。结论特异性DcR3-siRNA在体外实验能抑制目的基因的表达。

Inhibition of the expression of DcR3 in human hepatocellular carcinoma cell line HepG2

Objective To explore the effect of specific DcR3-siRNAs on the expression of DcR3 in human hepatocellular carcinoma cell line HepG2.Methods Designed and synthesised four specific siRNA targeting DcR3 and one random siRNA labeled with FAM.siRNAs were transfected into HepG2 cells by Lipofectamine TM2000,the transfection efficiency was detected by fluorescence microscope and the flow cytometry.Semi-quantitive RT-PCR and immunocyto-chemistry method was used to assess the change of the expression of DcR3.Results The four specific siRNAs targeting DcR3 all decreased the expression of DcR3 mRNA.The best siRNA was siRNA4,and it could silence the mRNA expression with the silencing rate of 62.9%.The expression of DcR3 protein could be inhibited by siRNA4.Conclusion Specific siRNA targeting DcR3 can inhibit the expression of DcR3.