首页|(-)-表儿茶素通过调控A549R细胞自噬改善放疗敏感性的作用机制

(-)-表儿茶素通过调控A549R细胞自噬改善放疗敏感性的作用机制

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目的 探讨(-)-表儿茶素[(-)-Epicatechin,EC]通过联合放疗(Radiotherapy,RT)改善非小细胞肺癌细胞放疗敏感性的作用机制.方法 体外培养A549人非小细胞肺癌细胞,构建放疗抵抗株A549R.通过细胞计数试剂盒8(Cell Counting Kit-8,CCK-8)检测细胞增殖活力,脱氧核苷酸末端转移酶介导的dUTP缺口末端标记(terminal deoxy-nucleotidyl transferase-mediated dUTP nick-end labeling,TUNEL)染色观察细胞凋亡情况,Transwell 实验检测细胞迁移能力.蛋白质免疫印迹(Western blot)检测凋亡相关蛋白(Bcl-2、Bax、Cleaved Caspase-3)和自噬相关蛋白(Beclin、LC3Ⅱ/Ⅰ)的表达.免疫荧光染色检测Cleaved Caspase-3和LC3的表达.单丹磺酰尸胺(Monodansylcadaverine,MDC)染色检测自噬体的形成.结果 EC联合RT抑制细胞增殖和迁移能力、诱导细胞凋亡和自噬、下调Bcl-2的表达、上调Bax、Cleaved Caspase-3、Beclin和LC3 Ⅱ/Ⅰ的表达,而加入自噬抑制剂(Chloroquine,CQ)或mTOR信号通路激活剂MHY1485处理后部分减弱了 EC联合RT处理对细胞的作用.结论 EC可作为放疗增敏剂,通过抑制mTOR信号通路诱导A549R细胞自噬和凋亡改善放疗敏感性.
Mechanism of EC Regulation of Autophagy in A549R Cells to Improve Radiotherapy Sensitivity
Objective To investigate the mechanism of(-)-epicatechin(EC)in improving the radiosensitivity of non-small cell lung cancer cells by combining radiotherapy(RT).Methods A549 human non-small cell lung cancer cells were cul-tured in vitro to construct a radiotherapy-resistant cell line A549R.Cell proliferation activity was detected by Cell Counting Kit-8(CCK-8).The cell apoptosis was observed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labe-ling(TUNEL)staining.Transwell assay was used to detect cell migration ability.The expressions of apoptosis-related proteins(Bcl-2,Bax,Cleaved Caspase-3)and autophagy-related proteins(Beclin,LC3Ⅱ/Ⅰ)were detected by Western blot.The ex-pressions of Cleaved Caspase-3 and LC3 were detected by immunofluorescence staining.Monodansylcadaverine(MDC)staining was used to detect the formation of autophagosomes.Results EC combined with RT inhibited cell proliferation and migration,in-duced cell apoptosis and autophagy,down-regulated the expression of Bcl-2,up-regulated the expressions of Bax,Cleaved Caspase-3,Beclin and LC3Ⅱ/Ⅰ,while the effect of EC combined with RT was partially attenuated by the treatment with autoph-agy inhibitor(chloroquine,CQ)or mTOR signaling pathway activator MHY1485.Conclusion EC can be used as a radiosensitizer to improve the radiosensitivity of A549R cells by inhibiting mTOR signaling pathway to induce autophagy and apoptosis.

non-small cell lung cancer(-)-epicatechinradiotherapy sensitizerautophagymTOR signaling pathway

黄梅芳、王俊峰、岳军、李田芊、繆怡、王淋、何永梅、康敏、母丽霞、李文辉

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曲靖市第一人民医院,云南曲靖 655000

非小细胞肺癌 (-)-表儿茶素 放疗增敏剂 自噬 mTOR信号通路

国家自然科学基金云南省基础研究专项重点项目云南省高层次人才培养支持计划"名医"专项云南省高层次人才培养支持计划"名医"专项

82160470202001AS070011云厅字[2018]11号YNWR-MY-2020-016

2024

10.13193/j.issn.1673-7717.2024.03.009

中华中医药学刊
中华中医药学会 ,辽宁中医药大学

中华中医药学刊

CSTPCD北大核心
影响因子：1.007
ISSN：1673-7717
年,卷(期)：2024.42(3)
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