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目的研究RNA干扰(RNAi)对兔血管平滑肌细胞(VSMCs)bcl-2基因表达的干扰效应。方法用脂质体法将构建的装载靶向bcl-2基因小干扰性RNA(siRNA)的表达载体(pshRNA-bcl-2质粒)转染VSMCs(基因组),以转染空载体的细胞和加DMEM的空白细胞作为对照,采用半定量RT-PCR和Westernblot法检测bcl-2基因的表达,用MTT法检测VSMCs生长情况。结果转染siRNA的表达载体可以抑制内源性bcl-2基因在转录和转译水平上的表达,基因组bcl-2的mRNA和蛋白表达较空载体组和空白对照组明显减少(P<0.01);基因组的VSMCs生长也较空载体组和空白对照组明显受到抑制(P<0.01)。结论载体介导的RNAi技术可明显抑制内源性bcl-2基因的表达和VSMCs生长。
Objective To investigate the interference effect of RNAi on bcl-2 gene expression in rabbit vascular smooth muscle cells (VSMCs). Methods The constructed expression vector (pshRNA-bcl-2 plasmid) containing small interfering RNA (siRNA) targeting the bcl-2 gene was transfected into VSMCs (genomes) by liposome method, and transfected into empty vector cells The blank cells of DMEM served as controls. The expression of bcl-2 gene was detected by semi-quantitative RT-PCR and Western blot. The growth of VSMCs was detected by MTT assay. Results The expression of bcl-2 gene was inhibited by transfection with siRNA. The mRNA and protein expression of bcl-2 mRNA and protein were significantly decreased (P <0.01) compared with the blank vector group and the blank control group. The growth of VSMCs in the genome was also significantly inhibited compared with the empty vector group and the blank control group (P <0.01). Conclusion Vector-mediated RNAi can significantly inhibit the expression of endogenous bcl-2 gene and the growth of VSMCs.