松萝酸对A549和NCI-H358细胞增殖、凋亡及自噬的影响  

Effects of usnic acid on proliferation,apoptosis and autophagy of A549 and NCI-H358 cells

作　　者：李新伟 王延婷 李瑞雪 白慧敏[1] 刘锦龙[1] Li Xinwei;Wang Yanting;Li Ruixue;Bai Huimin;Liu Jinlong(Teaching and Research Section of Immunology,Baotou Medical College,Baotou 014040)

机构地区：[1]包头医学院免疫学教研室,包头014040

出　　处：《安徽医科大学学报》2025年第3期455-462,共8页Acta Universitatis Medicinalis Anhui

基　　金：内蒙古自治区自然科学基金项目(编号:2020LH08006)。

摘　　要：目的 探究松萝酸(UA)对肺癌A549和NCI-H358细胞增殖、细胞周期、凋亡和自噬的影响。方法 CCK-8法检测UA对两种肺癌细胞增殖抑制作用。流式细胞术检测UA对两种肺癌细胞周期阻滞作用。DCFH-DA探针法和流式细胞术检测UA诱导两种肺癌细胞生成活性氧(ROS)。Western blot检测两种肺癌细胞经UA及UA+ROS抑制处理后凋亡相关蛋白Bax、Caspase-3和Cleaved-Caspase-3及自噬相关蛋白LC3-Ⅰ和LC3-Ⅱ的表达情况。结果 (1) UA降低了两种细胞存活率,且抑制A549细胞增殖能力强于NCI-H358细胞。(2) UA将A549细胞阻滞在G0/G1期,NCI-H358细胞阻滞在G2/M期和S期。(3) UA诱导两种细胞ROS含量增加,NCI-H358细胞ROS增加量多于A549细胞,ROS抑制剂会减少UA诱导的细胞内ROS增多。(4) UA引起了两种肺癌细胞凋亡相关蛋白Bax和Cleaved-caspase-3高表达及自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ比值升高,其对A549促凋亡和自噬作用强于NCI-H358;抑制ROS后,两种肺癌细胞的Bax和Cleaved-Caspase-3表达量和LC3-Ⅱ/LC3-Ⅰ比值均降低,其中NCI-H358细胞降低更多。结论 UA抑制肺癌A549和NCI-H358细胞增殖,诱导细胞周期阻滞,并引起癌细胞发生细胞凋亡与自噬,且UA对A549细胞抑制杀伤作用强于NCI-H358细胞。这其中诱生细胞ROS参与了UA对两种肺癌细胞的作用,相比A549细胞,ROS可能更多介导UA诱导NCI-H358细胞凋亡和自噬发生。Objective To investigate the effects of usnic acid(UA)on proliferation,cell cycle,apoptosis and autophagy of lung cancer cells A549 and NCI-H358.Methods The CCK-8 method was used to detect the inhibitory effect of UA on two kinds of lung cancer cells proliferation.Flow cytometry was used to detect the cell cycle arrest effect of UA on two types of lung cancer cells.The fluorescence amount of UA-induced reactive oxygen species(ROS)in two kinds of lung cancer cells were detected by DCFH-DA probe assay and flow cytometry.Western blot was used to detect the expression of apoptosis related proteins Bax,Caspase-3,Cleaved-Caspase-3,and autophagy related proteins LC3-Ⅰand LC3-Ⅱin two types of lung cancer cells after treatment with UA and UA+ROS inhibition.Results①Usnic acid reduced the survival rates of two types of cells and had a stronger ability to inhibit the proliferation of A549 cells than NCI-H358 cells.②Usnic acid blocked A549 cells in the G 0/G 1 phase,while NCI-H358 cells in the G 2/M and S phases.③Usnic acid induced an increase in ROS content in two types of cells.Compared to A549,NCI-H358 cells showed a greater increase in ROS,and the ROS inhibitor reduced the intracellular ROS increase induced by UA.④Usnic acid induced high expression of apoptosis-related proteins Bax and Cleaved Caspase-3 and increased the ratio of autophagy-related proteins LC3-Ⅱ/LC3-Ⅰin both lung adenocarcinoma cells,and its pro-apoptotic and autophagic effects were stronger in A549 than in NCI-H358.After ROS inhibition,the expression levels of Bax and Cleaved Caspase-3 and the LC3-Ⅱ/LC3-Ⅰratio of both lung adenocarcinoma cells decreased,and the decrease was greater in NCI-H358 cells.Conclusion Usnic acid inhibits the proliferation of lung cancer A549 and NCI-H358 cells,induces cell cycle arrest,and induces apoptosis and autophagy in cancer cells.The inhibitory and killing effect of UA on A549 cells is stronger than that on NCI-H358 cells.In this case,the induced cell ROS are involved in the action of UA in two types o

关 键 词：UA A549 NCI-H358 细胞周期 活性氧 凋亡 自噬 

分 类 号：R392[医药卫生—免疫学]