蛋白酶体抑制剂诱导卵巢癌SKOV3细胞凋亡中伴随caspase依赖的BAG3剪切

Caspase-dependent cleavage of BAG3 in proteasome inhibitors-induced apoptosis in SKOV3 ovarian cancer cells

目的:探讨Bcl-2相关抗凋亡基因3(Bcl-2-associated athanogene 2,BAG3)在蛋白酶体抑制剂诱导卵巢癌SKOV3细胞凋亡中的作用。方法:选取人卵巢癌细胞系SKOV3,分别设培养液处理空白对照组、蛋白酶体抑制剂MG132处理组和MG132+Z-VAD联合处理组,按不同时段处理细胞。实时定量RT-PCR法检测各组细胞BAG3 mRNA表达及MG132诱导其表达时效性,Western blot蛋白印迹法检测各组细胞BAG3蛋白表达,流式细胞仪(FCM)检测细胞凋亡。结果:与空白对照组相比,在MG132诱导作用下,SKOV3细胞中BAG3 mRNA表达增加(P<0.01),并随作用时间呈现时效性。发现40kDa BAG3分裂产物,其细胞凋亡率亦随之增加。结论:蛋白酶体抑制剂可诱导上调卵巢癌细胞中BAG3基因表达,BAG3蛋白裂解导致细胞凋亡增加,这一效应可能依赖caspase介导完成。

Objective：To investigate the role of BAG3 in proteasome inhibitor mediated apoptosis in ovarian cancer cells. Methods： SKOV3 cell was cultured and treated with vehicle, MG132, pancaspase inhibitor Z - VAD or combination of Z -VAD and MG132. BAG3 mRNA and protein levels were analyzed by using qPCR and Western blot,respectively. Flow cytometry （FCM） was used to determine the apoptotic rate of cells. Results： Compared with the blank group,BAG3 mRNA expression in the ceils was increased by MG132 in a time -dependence manner （P 〈0. 01 ） ,and simultaneously,cleavage product of BAG3 could be induced by MG132, accompanied with increasing apoptosis rate. Conclusion：Proteasome inhibitor MG132 induces the expression of BAG3 in SKOV3 cells. In proteasome inhibitor treated ceils, BAG3 is cleaved in a caspase- dependent manner.