顺铂诱导肾小管上皮细胞毒性及Bcl-2对抗细胞凋亡作用机理研究

目的探讨顺铂诱导肾小管上皮细胞(RPTC)凋亡的作用机制和BCL-2基因抑制顺铂诱导细胞凋亡的作用途径。方法应用不同亚细胞结构分布的BCL-2基因体外转染RPTC,激光共焦显微技术和免疫荧光技术分析BCL-2(X)基因表达产物在RPTC线粒体和内质网上的分布定位,采用Hoechst33258染色并进行细胞凋亡计数。结果不同畸变的BCL-2(BCL-acta,BCL-cb5)分别定位于RPTC的线粒体、内质网,BCL-nt同时定位于上述两种细胞器。BCL-cb5组可见更多BAX被激活,同时可见较多碎裂细胞核。定位于线粒体上的BCL-acta明显抑制顺铂对RPTC诱导的细胞凋亡(P<0.05)。结论BCL-2抑制顺铂诱导RPTC凋亡主要是通过线粒体途径起作用。

RPTC cells toxicity induced by cisplatin and anti-apoptosis mechanism of BCL-2

Objective To explore the mechnism of toxicity induced by cisplatin on renal proximal tubular cells(RPTC) line and anti-apoptosis mechanism of BCL-2 with transfection of different mutant BCL-2 in vitro.MethodsRPTC was translocated with different mutants BCL-2(s).Cell apoptosis induced by cisplatin on RPTC was analyzed with confocal and flurencent microscope.The cell apoptosis was measured with Hoechst33258 after treatment with cisplatin.Results Different mutant BCL-2(BCL-acta,BCL-cb5)were translocated on mitochondrial and Endoplasmic Reticulum(ER) respectively.BCL-acta protected RPTC from apoptosis induced by cisplatin more easily than BCL-cb5 group in a time-dependant manner(P<0.05).Conclusion Cisplatin can induce RPTC apoptosis obivously through a mitochondrial signaling payhway and BCL-2 can protect more RPTC from apoptosis in the mitochondrial manner other than in ER manner