3－甲基腺嘌呤对喜树碱诱导的宫颈癌Hela细胞凋亡的影响

摘要

Objective:To explore the effect of autophagy inhibitor 3-methyladenine(3-MA) on camptothecin(CPT)-induced Hela cell apoptosis.Methods:MTT assays were carried out to determine the optimal concentration and time of CPT on Hela cells and the effect of different drugs on Hela cell proliferation activity .After Hela cells were treated with different drugs ,the changes of autophagy marker protein( microtubule-associated protein 1 light chain 3,LC3),p62 and apoptosis-related protein were detected using Western blot and immunofluorescence ( IF) .DAPI ( nuclear ) staining was used to observe cell apoptosis rate .Results: In CPC-treated Hela cells,Hela cell proliferation activity declined dramatically ,and autophagy could be induced to occur .Compared with CPT group ,the cell proliferation activity was lower in CPT combined with 3-MA group,the level of autophagy decreased ,but the apoptosis rate significantly increased.Conclusion:CPT can induce autophagy while inducing Hela cell death .Hela cells chemosensitivity to CPT treatment can be enhanced by 3-MA inhibiting autophagy .%目的：本研究探讨自噬抑制剂3－甲基腺嘌呤（3－methyladenine，3－MA）对喜树碱（Camptothecin，CPT）诱导的Hela细胞凋亡的影响。方法：用四甲基偶氮唑盐比色法（ MTT方法）检测CPT对Hela细胞作用的最佳药物浓度和时间，以及不同药物对Hela细胞增殖活性的影响；用免疫印迹及免疫荧光检测不同药物作用于Hela细胞后，Hela细胞自噬标志蛋白微管相关蛋白1轻链3（microtubule－associated protein 1 light chain 3，LC3）、p62及凋亡相关蛋白的变化；DAPI核染色观察细胞凋亡。结果：CPT作用于Hela细胞后，Hela细胞增殖活性明显下降，并且可诱导自噬现象的发生。 CPT和3－MA联合作用较单独CPT作用Hela细胞的增殖活性降低，细胞自噬水平下降，凋亡率明显升高。结论：CPT在诱导Hela细胞凋亡的同时可诱导自噬，通过3－MA抑制自噬可增强Hela细胞对CPT作用的敏感性。