Construction of short hairpin RNA expression plasmid and its inhibition of survivin expression
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摘要:
目的： 探讨survivin短发夹RNA(shRNA)的表达质粒对胆囊癌细胞survivin mRNA及蛋白表达的抑制作用。方法：构建pshRNA-survivin重组质粒，在脂质体的介导下转染胆囊癌细胞株GBC-SD。RT-PCR分析survivin mRNA的表达;Western blot 检测survivin蛋白表达。结果：PCR和DNA测序证实表达质粒构建成功，并能明显地抑制GBC-SD/survivin mRNA的表达。结论：构建的pshRNA-survivin表达质粒能有效地抑制转染细胞survivin mRNA的表达，从而为肿瘤的生物学治疗提供新的方法和材料。
Abstract:
Abstract:Objective：To study the inhibit effect of survivin short hairpin RNA on the survivin mRNA and protein expression of gallbladder carcinoma cells. Methods：Human gallbladder carcinoma cells(GBC-SD) were transfected with recombinant plasmid. RT-PCR and Western blot were used to analyze the changes in expression levels of survivin mRNA and protein. Results：The size of the PCR product was 350bp. DNA sequencing showed that the sequence of recombinant vector pshRNA-survivin was successfully constructed and suppressed the expression of (GBC-SD) survivin mRNA. Conclusions：The recombinant plasmid constructed can inhibit the expression of survivin mRNA in transfected cells. This provides a new method and material for the biological therapy of cancer.