vegf shRNA 对耐药白血病细胞化疗敏感性的增强作用

本研究采用RNA干扰技术探讨血管内皮生长因子(VEGF)对耐药白血病细胞K562/A02药物敏感性的影响及其机制。针对人vegf基因合成3个特异性shRNA干扰片段,采用脂质体介导的方法将其导入K562/A02细胞,用RT-PCR检测vegf及mrp1的mRNA表达水平,用Western blot检测VEGF、MRP1、AKT及P-AKT的蛋白表达情况,用MTT法检测阿霉素对各组细胞的半数抑制浓度(IC50),用流式细胞术检测细胞凋亡、胞内罗丹明123(Rho123)积聚情况。结果表明:转染vegf shRNA后,K562/A02细胞vegf的mRNA表达下调,其中vegf shRNA2组和vegf shRNA3组与HK组比较有显著差异(p<0.05),以vegf shRNA3组最显著,VEGF蛋白表达也出现下调,与mRNA的表达基本一致;MTT检测结果显示阳性转染组对阿霉素的敏感性增加,其中vegf shRNA2组和vegfshRNA3组的半数抑制浓度(IC50)与HK组比较具有显著的统计学差异(p<0.05);阳性转染组胞内Rho123积聚增多,与HK组比较均有统计学差异(p<0.05);阳性转染组细胞凋...

vegf shRNA Enhances the Sensitivity of Multidrug-resistant Leukemia Cells to Anticancer Agent

This study was aimed to explore the effect of vascular endothelial growth factor(VEGF) on sensitivity of leukemia cell line K562/A02 to doxorubicin by using RNA interference,and to investigate its mechanism.The 3 shRNA targeting human vegf gene were synthesized,then transfected into K562/A02 cells by lipofectamine 2000 reagent.RT-PCR was used to detect the expression of vegf and mrp1 at the mRNA level;Western blot was used to analyze the expression of VEGF,MRP1,AKT,P-AKT at the protein level;MTT was used to...