Survivin反义核酸诱导肝癌细胞凋亡及降低化疗药物耐药的研究

Study on apoptosis of liver cancer cells induced by Survivin antisense RNA and reduce the chemotherapy drug resistance

目的构建Survivin反义核酸真核表达载体(anti-pcDNA3-svv),探讨其对肝癌细胞的凋亡诱导作用及降低化疗耐药的机制。方法采用Survivin反义核酸瞬时电转染法转染肝癌细胞系Skov3,筛选出阳性细胞株。以Western-blot检测生存素蛋白的表达,用流式细胞仪检测其对Skov3的凋亡诱导作用,并检测实验组细胞(Skov3-SVVanti)MDR-1mRNA的表达。结果稳定表达an-ti-pcDNA3-svv的Skov3-SVVanti其生存素蛋白表达比未转染者明显降低,Skov3-SVVanti组细胞流式细胞仪检测显示其凋亡率为19%。Skov3-SVVanti组MDR-1mRNA表达较Skov3组明显减少(P<0.01)。结论 Survivin反义核酸可诱导肝癌细胞凋亡,并通过降低MDR-1mRNA表达,降低化疗药物耐药性。

Objective To establish Survivin antisense RNA and to explore its effect on the apoptosis of liver cancer cells and the mechanism of reduce the chemotherapy drug resistance.Methods Survivin antisense eukaryotic vector anti-pcDNA3-svv was transferred into Skov3 cells by electroporation and positive clone was screened out.Survivin protein was detected by Western blot;effect of apoptosis inducement was observed by flow cytometry;expression of MDR-1 mRNA was detected.Results Survivin protein of positive clone cells reduced obviously compared with non-transfered cells.Flow cytometry showed the apoptosis rate in Skov3-SVVanti group was 19%.Expression of MDR-1 mRNA in Skov3-SVVanti group reduced obviously compared with Skovg 3 group.Conclusion Surivivin antisense RNA can induce liver cancer cell apoptosis and reduce the resistance of chemotherapy drug by reducing expression of MDR-1 mRNA.