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目的探讨Livin-siRNA对人乳腺癌细胞株MCF-7和ZR-75-30中Livin基因及蛋白表达的下调作用,以及对其增殖能力的影响。方法构建3个靶向Livin的siRNA真核表达载体,分别转染乳腺癌细胞后,通过实时定量PCR、Western blot技术检测人乳腺癌细胞MCF-7,ZR-75-30的Livin在mRNA、蛋白水平的表达。采用MTT法检测si-Livin2对乳腺癌细胞增殖的抑制作用。结果成功构建了针对Livin基因的3个特异性siRNA表达载体。它们不同程度地抑制了乳腺癌细胞Livin基因的表达,以si-Livin2抑制效率最高。si-Livin2作用于人乳腺癌细胞株MCF-7和ZR-75-30后能明显抑制其Livin基因的mRNA及蛋白表达水平(P<0.05),并且能有效抑制细胞的增殖。结论Livin-siRNA能特异性抑制乳腺癌细胞的Livin mRNA及其蛋白的表达,抑制细胞增殖,为进一步研究沉默Livin基因对人乳腺癌细胞生物学功能的影响奠定了基础。
Objective To investigate the down-regulation of Livin gene and protein expression in human breast cancer cell lines MCF-7 and ZR-75-30 by Livin-siRNA and its effect on proliferation. Methods Three siRNA targeting Livin siRNA eukaryotic expression vectors were constructed and transfected into breast cancer cells respectively. The mRNA and protein expression of Livin in human breast cancer cells MCF-7 and ZR-75-30 were detected by real-time PCR and Western blot, Horizontal expression. The inhibitory effect of si-Livin2 on the proliferation of breast cancer cells was detected by MTT assay. Results Three specific siRNA expression vectors targeting Livin gene were successfully constructed. They inhibit the expression of Livin gene in breast cancer cells to varying degrees, with the highest inhibition rate of si-Livin2. The effect of si-Livin2 on human breast cancer cell lines MCF-7 and ZR-75-30 significantly inhibited the mRNA and protein expression of Livin gene (P <0.05) and inhibited cell proliferation effectively. Conclusion Livin-siRNA can specifically inhibit Livin mRNA and protein expression and inhibit cell proliferation in breast cancer cells, which lays the foundation for further study of the effect of silencing Livin gene on the biological function of human breast cancer cells.