沉默CtBP1基因逆转人子宫内膜癌耐药细胞株B-MD-C1(ADR^(+/+))的多药耐药性

Silencing CtBP1 gene reverses the multidrug resistance of drug-resistant endometrial cancer B-MD-C1(ADR^(+/+)) cells

目的:探讨短发夹式RNA(short hairpin RNA,shRNA)干扰羧基末端结合蛋白1(C-terminal-binding proteins 1,CtBP1)基因表达是否可以逆转多药耐药基因1(multidrug resistant gene 1,MDR1)介导的人子宫内膜癌耐药细胞株B-MD-C1(ADR+/+)的多药耐药性。方法:采用脂质体转染法将特异性的shRNA1和shRNA2及非特异性的shRNAHK质粒转染入人子宫内膜癌耐药细胞株B-MD-C1(ADR+/+)中,RT-PCR法检测各组细胞MDR1基因表达水平,Western印迹法检测各组细胞P糖蛋白(P-glycoprotein,P-gp)的表达,MTT法测定各组细胞对多柔比星的敏感性变化。结果:RT-PCR结果显示,shRNA1组和shRNA2组细胞中MDR1基因被抑制,基因表达抑制率约50%;Western印迹结果显示,shRNA1组和shRNA2组细胞P-gp蛋白的表达水平显著降低;MTT检测发现,shRNA转染48和72h时shRNA1组和shRNA2组细胞对多柔比星的敏感性增加(P<0.01)。结论:沉默CtBP1基因可抑制人子宫内膜癌耐药细胞株B-MD-C1(ADR+/+)中MDR1基因和P-gp蛋白的表达,降低细胞的耐药性。

Objective：To study whether the short hairpin RNA （shRNA） targeting C-terminal binding proteins 1 （CtBP1）gene could reverse the multidrug resistance of endometrial cancer B-MD-C1 （ ADR^＋/＋ ） induced by muhidrug resistant gene 1 （MDR1）. Methods ： The plasmid expression vectors of shRNA1, shRNA2 and non-specific shRNAHK were transfected into human endometrial cancer B-MD-C1 （ ADR^ ＋/＋ ） cells with LipofectamineTM2000. The expression of MDR1 gene was detected with RT-PCR and expression of P-glycoprotein （P-gp） was determined with Western blotting. The sensitivity of cells to adriamycin was evaluated by MTT assay. Results： RT-PCR demonstrated that the expression of MDR1 gene was inhibited in shRNAt and shRNA2 groups, and the inhibitory rate was about 50%. Western blotting showed that the expression of P-gp was significantly decreased in B-MD-C1 （ADR ^＋/＋ ） ceils transfected with shRNAt or shRNA2. The sensitivity of B-MD-C1 （ ADR ^＋/＋ ） cells to adriamycin was increased at 48 and 72 h after transfection of shRNAI or shRNA2 （P 〈 0.01 ）. Conclusion：Silencing CtBP1 gene expression inhibits the expressions of MDR1 gene and P-gp and decreases the muhidrug resistance of human endometrial cancer B-MD-C1 （ADR^＋/＋）cell.