论文部分内容阅读
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。
Objective To investigate the mechanism of cisplatin-induced apoptosis of renal tubular epithelial cells (RPTC) and the role of BCL-2 gene in cisplatin-induced apoptosis. Methods BCL-2 gene was transfected into RPTC by laser scanning confocal microscopy and immunofluorescence technique. The distribution of BCL-2 (X) gene in mitochondria and endoplasmic reticulum of RPTC was analyzed by Hoechst33258 staining Apoptosis was counted. Results BCL-2 (BCL-acta, BCL-cb5) with different aberrations were localized in the mitochondria, endoplasmic reticulum and BCL-nt of RPTC, respectively, in the two organelles. In the BCL-cb5 group, more BAXs were activated with more fragmented nuclei. BCL-acta located on mitochondria significantly inhibited cisplatin-induced apoptosis (P <0.05). CONCLUSION BCL-2 inhibits cisplatin-induced apoptosis in RPTC primarily through mitochondrial pathway.