论文部分内容阅读
目的采用自噬特异性抑制剂3-甲基腺嘌呤(3-MA)抑制自噬,观察其对顺铂(DDP)诱导HeLa细胞凋亡的影响,探讨自噬在DDP诱导人宫颈癌HeLa细胞凋亡中的作用。方法用四甲基偶氮唑盐比色法(MTT)观察3-MA对DDP抑制HeLa细胞生长情况的影响;倒置相差显微镜观察细胞形态变化;间接免疫荧光法检测自噬标志蛋白LC3和p62的变化;Hoechst荧光染色检测细胞凋亡。结果 MTT法显示,3-MA与DDP联合应用导致HeLa细胞生长抑制率增加;光镜结果显示,3-MA与DDP联合应用加剧了HeLa细胞收缩变圆,促进死亡;间接免疫荧光法检测到3-MA与DDP联合应用减弱了DDP诱导的HeLa细胞LC3聚集及其与p62的共定位;Hoechst荧光染色检测到3-MA与DDP联合应用增加HeLa细胞凋亡率。结论 3-MA抑制自噬能够促进DDP诱导HeLa细胞发生凋亡。
OBJECTIVE: To investigate the effect of autophagy on the apoptosis of HeLa cells induced by cisplatin (DDP) and to explore the possible mechanism of autophagy in DDP-induced human cervical carcinoma HeLa cells The role of apoptosis. Methods MTT assay was used to observe the effects of 3-MA on HeLa cell proliferation induced by DDP. Morphological changes were observed by inverted phase contrast microscope. Indirect immunofluorescence assay was used to detect the expression of autophagy markers LC3 and p62 Changes; Hoechst fluorescence staining for apoptosis. Results MTT assay showed that the combination of 3-MA and DDP led to an increase in the growth inhibition rate of HeLa cells. The results of light microscopy showed that the combination of 3-MA and DDP exacerbated the contraction of HeLa cells and promoted death; indirect immunofluorescence assay detected 3 The combination of -MA and DDP attenuated the DLC-induced LC3 accumulation in HeLa cells and colocalized with p62. Hoechst staining detected that the combination of 3-MA and DDP increased the apoptosis rate of HeLa cells. Conclusion 3-MA inhibition of autophagy can promote apoptosis induced by DDP in HeLa cells.