【摘要】 目的:研究JWA基因(又称ARL6IP5基因,ADP-ribosylation-like factor 6 interacting protein 5)对肿瘤细胞P-糖蛋白表达及其功能的影响。方法:利用脂质体转染技术将JWA-shRNA重组质粒及其空载质粒转入人绒毛膜癌细胞JAR和人乳腺癌细胞MCF-7,将Flag-JWA重组质粒及其空载质粒转入人绒毛膜癌耐依托泊苷(etoposide,VP16)JAR/VP16细胞;Westernblotting检测JWA和P-糖蛋白的表达,流式细胞术分析细胞内罗丹明123(Rhodamine 123,Rh123)的潴留情况。结果:JWA-shRNA重组质粒转入JAR细胞和MCF-7细胞后,JWA表达水平降低,P-糖蛋白表达水平上调,罗丹明潴留减少;Flag-JWA重组质粒转入JAR/VP16细胞后,JWA表达水平上调,P-糖蛋白表达水平下降,罗丹明潴留增加。结论:JWA基因可以调控肿瘤细胞P-糖蛋白的表达,并影响其转运功能。更多还原

【Abstract】 Objective To study the effects of JWA(ADP-ribosylation-like factor 6 interacting protein 5,ARL6IP5) gene on expression and function of P-glycoprotein in tumor cells.Methods: JWA-shRNA and control-shRNA plasmids were transfected into human choriocarcinoma cell line JAR and human breast cancer cell line MCF-7,and flag-JWA and flag-control plasmids were transfected into etoposide(VP16)-resistant JAR cells(JAR/VP16) by liposome-mediated transfection assay.JWA and P-glycoprotein expressions were examined by Western blotting analysis.Intercellular retention of rhodamine 123(Rh123) was determined by FCM.Results: After JWA expression was down-regulated by JWA-shRNA transfection,P-glycoprotein expression was increased in JAR and MCF-7 cells,and Rh123 retention was decreased.After JWA was over-expressed by flag-JWA transfection,P-glycoprotein expression was decreased in JAR /VP16 cells and Rh123 retention was increased.Conclusion: JWA gene can regulate the expression and transportation of P-glycoprotein in tumor cells.更多还原