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

doi:10.3969/j.issn.1004-616x.2024.01.002

摘要/Abstract

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。

关键词: 自噬, α粒子, 人支气管上皮细胞, 癌变, 肺癌

Abstract: 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 60 μmol/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 60 μmol/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 for48 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.

Key words: autophagy, alpha particle, human bronchial epithelial cells, canceration, lung cancer

中图分类号:

R734.1

杨莉, 邵帅, 武云云, 王成芳, 曲功霖, 闫皓宇, 苟巧. 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[J]. 癌变·畸变·突变, 2024, 36(1): 9-15,20.

YANG Li, SHAO Shuai, WU Yunyun, WANG Chengfang, QU Gonglin, YAN Haoyu, GOU Qiao. Role of autophagy in alpha particle-induced malignant transformation of human bronchial epithelial cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(1): 9-15,20.

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