细胞自噬减缓PM2.5诱导的人肺腺癌细胞系H441凋亡  

Cell autophagy attenuates PM2.5-induced apoptosis of human lung adenocarcinoma cell line H441

作　　者：李春艳 王亚红 陈婷 杨拉维 刘刚 LI Chun-yan;WANG Ya-hong;CHEN Ting;YANG La-wei;LIU Gang(Dept.of Respiratory Medicine,the Affiliated Hospital of Guangdong Medical University,Zhanjiang 524001,China;Clinical Research Center,the Affiliated Hospital of Guangdong Medical University,Zhanjiang 524001,China)

机构地区：[1]广东医科大学附属医院呼吸内科,广东湛江524001 [2]广东医科大学附属医院临床医学研究中心,广东湛江524001

出　　处：《基础医学与临床》2018年第7期907-912,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81570062);广东省医学科研基金(A2017027);广东医科大学科研基金(M2016001)

摘　　要：目的探讨细胞自噬在PM2.5诱导细胞凋亡过程中的作用机制。方法采集2016年湛江市细颗粒物PM2.5,再用不同浓度PM2.5,分不同时间处理人肺腺癌(H441)细胞;用MTT比色法检测细胞增殖;PI和annexin V双染及TUNEL法检测细胞凋亡;蛋白免疫印迹法检测自噬标志物LC3Ⅱ及AKT和P-AKT蛋白表达。将H441细胞用雷帕霉素或3-MA预处理后,再用PM2.5暴露处理,锥虫蓝染色检测细胞活力。结果与对照组相比,100μg/m L以上PM2.5处理24和48 h组细胞增殖明显受到抑制;随着PM2.5作用浓度的增加,细胞凋亡率明显增加,LC3Ⅱ蛋白的表达明显增多,P-AKT蛋白表达明显减少;而使用AKT抑制剂后,LC3Ⅱ蛋白增加愈明显;PM2.5暴露下,应用雷帕霉素能够降低PM2.5引起的细胞凋亡,而3-MA可促进PM2.5引起的细胞凋亡。结论在H441细胞中,PM2.5通过抑制AKT活化而激活细胞自噬发生,而细胞自噬能够减缓PM2.5诱导的细胞凋亡。Objective To investigate the function of autophagy in the process of PM2.5-induced apoptosis.Methods PM2.5 was obtained from Zhanjiang in 2016.Human lung adenocarcinoma cells H441 were treated with PM2.5 at different concentrations for different times.Cell proliferation was analyzed by MTT assay;Cell apoptosis was assessed by PI and Annexin V double staining and TUNEL assay.The expression of autophagy marker LC3Ⅱ,AKT and P-AKT protein was examined by Western blot(WB).H441 cells were treated with PM2.5 following treatment with rapamycin or 3-MA.Cell viability was evaluated by trypan blue staining.Results Compared with the control group,cell proliferation was significantly inhibited by PM2.5 at concentration of 100μg/mL or more for 24 and 48 h.With the increase of PM2.5 concentration,the cells apoptotic rate significantly increased,the protein expression of LC3Ⅱwas increased as well as the P-AKT was decreased;and the protein expression of LC3Ⅱwas increased significantly after AKT inhibitor treatment.Moreover,rapamycin decreased PM2.5-induced cell apoptosis,and 3-MA can promote PM2.5-induced cell apoptosis.Conclusions In H441 cells,PM2.5 activates autophagy by inhibiting activation of AKT pathway,and cell autophagy can mitigate PM2.5-induced apoptosis.

关 键 词：PM2.5 细胞自噬 AKT H441 

分 类 号：R562.2[医药卫生—呼吸系统]