自噬在α粒子辐射诱发人支气管上皮细胞恶性转化中的作用  

Role of autophagy in alpha particle-induced malignant transformation of human bronchial epithelial cells

作　　者：杨莉 邵帅[1] 武云云[1] 王成芳 曲功霖[1] 闫皓宇 苟巧[1] YANG Li;SHAO Shuai;WU Yunyun;WANG Chengfang;QU Gonglin;YAN Haoyu;GOU Qiao(Key Laboratory of Radiological Protection and Nuclear Emergency,National Institute for Radiological Protection,Chinese Center for Disease Control and Prevention,Beijing 100088,China)

机构地区：[1]中国疾病预防控制中心辐射防护与核安全医学所,辐射防护与核应急重点实验室,北京100088

出　　处：《癌变．畸变．突变》2024年第1期9-15,20,共8页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金(81000862)。

摘　　要：目的:探讨自噬在α粒子辐射诱发永生化人支气管上皮细胞BEP2D恶性转化中的作用。方法:采用Western blot检测BEP2D细胞、α粒子作用BEP2D后的第24代细胞RH24以及α粒子作用BEP2D后的恶性转化细胞BERP35T-1(经鉴定为肺鳞癌细胞)内自噬相关蛋白LC3B-II、LC3B-I和P62的表达,透射电镜观察细胞内单位面积自噬小体的数量。分别用40和60μmol/L自噬抑制剂氯喹(CQ)及25 pmol/L自噬激活剂雷帕霉素(Rapa)作用BERP35T-1细胞,采用Western blot法检测细胞内LC3B和P62蛋白的表达情况,CCK-8法检测细胞的存活率,Transwell侵袭实验检测细胞的侵袭数,划痕愈合实验检测细胞划痕闭合率。结果:与BEP2D细胞相比,RH24和BERP35T-1细胞内LC3B-II/I蛋白比值增高(P<0.05或P<0.01),P62蛋白表达降低,自噬小体数量增多(P<0.01)。40和60μmol/L的CQ分别作用BERP35T-1细胞48 h后,细胞内LC3B-II/I蛋白比值和P62蛋白表达水平均升高,细胞的存活率、侵袭数和划痕闭合率均降低(均为P<0.01);25 pmol/L的Rapa作用BERP35T-1细胞48 h后,细胞内LC3B-II/I蛋白比值升高,P62蛋白表达减弱,细胞的存活率、侵袭数和划痕闭合率均升高(P<0.05或P<0.01)。结论:在α粒子辐射诱发BEP2D细胞恶性转化过程中,细胞自噬增强,可能由此提高细胞的增殖、侵袭和迁移能力,从而促进细胞恶性转化。OBJECTIVE:To investigate the role of autophagy in malignant transformation of immortalized human bronchial epithelial cells BEP2D induced by alpha particle radiation.METHODS:Western blot was used to detect the expression of autophagy related proteins(LC3B-II,LC3B-I and P62)in alpha particles exposed BEP2D and in exposed malignant transformed cell line BERP35T-1.Intracellular autophagosomes were observed by transmission electron microscopy.BERP35T-1 cells were also treated with 40 and 60μmol/L autophagy inhibitor chloroquine(CQ)and 25 pmol/L autophagy activator Rapamycin(Rapa),respectively,for detection of of LC3Bs and P62.Survival rates of cells were measured by CCK-8,invasion ability by the Transwell invasion assay,and the migration ability by the scratch healing assay.RESULTS:Compared with BEP2D cells,the numbers of autophagosomes in RH24 and BERP35T-1 cells were increased,LC3B-II/I protein ratio increased,and P62 protein expression decreased.After treatment with 40 and 60μmol/L CQ for 48 h,the LC3B-II/I protein ratio and the expression level of P62 protein in BERP35T-1 cells were increased,and the cell survival rate,invasion number and scratch closure rate decreased(all P<0.01).After treatment with 25 pmol/L Rapa for 48 h,the LC3B-II/I protein ratios were increased,the expression of P62 protein decreased,and the survival rate of BERP35T-1 cells,invasion number and scratch closure rate were increased(P<0.05 or 0.01).CONCLUSION:During the alpha particle-induced malignant transformation of BEP2D cells,autophagy was enhanced which was associated with increased ability of cell proliferation,invasion and migration.

关 键 词：自噬 Α粒子 人支气管上皮细胞 癌变 肺癌 

分 类 号：R734.1[医药卫生—肿瘤]