Bcl-2基因siRNA的筛选及对胃癌细胞MGC-803增殖的影响作用

目的:筛选有效抑制Bcl-2基因表达的siRNA序列,研究Bcl-2特异性siRNA对胃癌细胞的抑制作用。方法:设计并合成4对针对Bcl-2的siRNA序列,通过LipofectamineTM2000转染入人胃癌细胞MGC-803后,应用实时定量PCR和Western Blot法分别检测siRNA对细胞内Bcl-2 mRNA和蛋白的表达水平抑制效果,且经CCK-8法检测siRNA对胃癌细胞增值影响。结果:与对照组相比,所设计4对siRNA中siBcl-2抑制Bcl-2 mRNA和蛋白表达效果最佳,抑制率分别为88%和63%,且siBcl-2在转染后第96 h对癌细胞增殖有显著抑制效果。结论:成功筛选出1对能有效抑制Bcl-2基因表达的siRNA序列,为胃癌基因治疗提供实验依据。

Screening of siRNA on Bcl-2 and Its Effect on Proliferation of Human Gastric Cancer Cell Line Cell MGC-803

Objective：To screen small interfering RNA（siRNA） sequences with Bcl-2 mRNA and study the inhibitory effect of Bcl-2-targeted siRNA in human MGC-803 gastric cancer cells.Methods：Four siRNAs targeting Bcl-2 gene were designed and synthesized,and they were transfeted into MGC-803 cells via LipofectimineTM2000.The inhibitory effect of siRNAs on the mRNA expression of Bcl-2 in infected cells were detected by real-time PCR,and on the protein exspression of Bcl-2 were tested by Western Blotting.The effect of suppressing proliferation of MGC-803 cells were detected by CCK-8.Results：Compared with the experimental and controls,siBcl-2,one of four couples of siRNAs,had the best inhibitory effect of Bcl-2 mRNA and protein expression,and the inhibition ratios were 88% and 63% respectively.The significant inhibitory effect of siBcl-2 on the reproduction of MGC-803 cells was observed as determined by CCK-8 after 96 hours transfection.Conclusion：The present experiment had successfully screened a couple of synthetic siRNA Sequence to Bcl-2 mRNA.It provided an useful experimental evidence for gene therapy of gastric cancer.