Wip1通过P-gp介导人多发性骨髓瘤细胞卡非佐米耐药作用研究

Wip1 mediating resistance to carfilzomib in human multiple myeloma cells via P-gp

目的 研究野生型p53诱导的蛋白磷酸酶1(Wip1)在多发性骨髓瘤细胞卡非佐米(CFZ)耐药中的作用及作用机制。方法 以多发性骨髓瘤细胞RPMI 8226作为研究对象,转染法过表达Wip1设为Wip1组,转染空载体设为NC组,同时以未转染细胞设为Control组;适时以CFZ处理细胞;MTT法检测细胞增殖能力;流式细胞术检测细胞凋亡;蛋白印记实验检测Wip1和P-糖蛋白(P-gp)蛋白表达;免疫荧光实验检测细胞P-gp蛋白表达。结果 与NC组相比,Wip1组Wip1蛋白表达显著上调(P<0.01)。与NC组相比,Wip1组在0.01~100μM CFZ处理后细胞增殖率均显著升高(P<0.01),CFZ对Wip1组细胞IC显著升高(P<0.01)。与NC+CFZ组相比,Wip1+CFZ组细胞凋亡显著降低(P<0.01)。与NC组相比,Wip1组P-gp蛋白表达显著上调(P<0.01)。结论 Wip1可能通过上调P-gp蛋白表达,诱导人多发性骨髓瘤细胞对卡非佐米耐药。

Objective To study the role and mechanism of p53-induced protein phosphatase 1(Wip1)in the resistance of multiple myeloma cells to carfilzomib(CFZ).Methods RPMI 8226 cells were used as the research subjects, the transfection method overexpressing Wip1 was set as Wip1 group, the transfected empty vector was set as NC group, and the untransfected cells were set as control group.The cells were treated with CFZ at the appropriate time.The cell proliferation ability was detected by MTT method;the cell apoptosis was detected by flow cytometry;Wip1 and P-glycoprotein(P-gp)protein expression was detected by Western blot.Immunofluorescence experiment was used to detect cell P-gp protein expression.Results Compared with NC group, the Wip1 protein expression in Wip1 group was significantly up-regulated(P<0.01).Compared with NC group, the cell proliferation rate of Wip1 group was significantly increased after 0.01~100 μM CFZ treatment(P<0.01),and the ICof the Wip group was significantly increased by CFZ(P<0.01).Compared with NC+CFZ group, cell apoptosis in Wip1+CFZ group was significantly reduced(P<0.01).Compared with NC group, the P-gp protein expression in Wip1 group was significantly up-regulated(P<0.01).Conclusion Wip1 can induce the resistance to carfilzomib in human multiple myeloma cells by up-regulating the expression of P-gp protein.