抑制自噬起始阶段增强喜树碱诱导的NCI-H1975人肺腺癌细胞凋亡  

Inhibition of autophagy initiation stage enhances camptothecin-induced apoptosis in NCI-H1975 cells

作　　者：张亚萍 曹丽 苏强[3] 程凯 张晓延 

机构地区：[1]山西医科大学汾阳学院医学检验系,山西汾阳032200 [2]山西医科大学 [3]山西医科大学附属第一医院神经内科重点实验室,山西太原030001

出　　处：《细胞与分子免疫学杂志》2017年第12期1640-1646,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金青年基金(81301426);山西医科大学校内重点项目(1208)

摘　　要：目的研究自噬抑制剂氯喹(CQ)和3-甲基腺嘌呤(3-MA)对喜树碱(CPT)诱导非小细胞肺腺癌NCI-H1975细胞凋亡的影响。方法通过CPT处理肺腺癌NCI-H1975细胞,CCK-8法检测细胞增殖活性,碘化丙啶(PI)染色观察细胞形态学的变化,流式细胞术检测CPT作用于NCI-H1975细胞后细胞的凋亡情况,Western blot法检测自噬及凋亡相关蛋白微管相关蛋白1轻链3Ⅰ(LC3Ⅰ)和LC3Ⅱ、P62、胱天蛋白酶3(caspase-3)和多腺苷二磷酸核糖聚合酶(PARP)、转化生长因子β1(TGF-β1)蛋白水平。结果 CPT处理后,LC3Ⅱ/LC3Ⅰ的比例增加;caspase-3和PARP发生明显降解;且这种降解作用能被3-MA增强而被CQ抑制。而且发现CPT处理后,引起细胞内TGF-β1降低。结论抑制NCI-H1975细胞自噬起始阶段后,增强CPT诱导细胞凋亡的敏感性。Objective To explore the effect of autophagic inhibitors chloroquine（ CQ） and 3-methyl adenine（ 3-MA） on apoptosis of non-small cell lung adenocarcinoma NCI-H1975 cells induced by camptothecin（ CPT）. Methods After NCI-H1975 cells were treated with CPT,cell proliferation was detected by CCK-8 assay,morphological changes of cells were observed by PI staining,and the apoptosis of NCI-H1975 cells was determined by flow cytometry. The levels of autophagyand apoptosis-related proteins LC3Ⅰ,LC3Ⅱ,P62,caspase-3 and poly ADP-ribose polymerase（ PARP） and transforming growth factor beta 1（ TGF-beta 1） were detected by Western blot analysis. Results After CPT treatment,the ratio of LC3Ⅱto LC3Ⅰ was raised. The apoptotic protease caspase-3 and substrate PARP were obviously degraded,which could be enhanced by 3-MA but inhibited by CQ. It was also found that the intracellular TGF-beta 1 was reduced after CPT treatment. Conclusion Inhibition of autophagy initiation stage in NCI-H1975 cells can increase the sensitivity of cell apoptosis induced by CPT.

关 键 词：喜树碱 NCI-H1975细胞 细胞凋亡 自噬 自噬抑制剂 

分 类 号：R392.12[医药卫生—免疫学] R965[医药卫生—基础医学]