miRNA-126通过抑制自噬逆转肺癌A549/DDP细胞顺铂耐药的作用研究  

miRNA-126 reverses cisplatin resistance in lung cancer A549/DDP cells by inhibiting autophagy

作　　者：唐夏莉 陈君 焦德敏 刘翔 TANG Xiali;CHEN Jun;JIAO Demin;LIU Xiang(The 903rd Hospital of PLA,Hangzhou 310004,China)

机构地区：[1]中国人民解放军联勤保障部队第九〇三医院,浙江杭州310004

出　　处：《中国病理生理杂志》2024年第8期1378-1383,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82101870);浙江省自然科学基金资助项目(No.LQ21H100001);浙江省中医药科技计划项目(No.2023ZL589);浙江省医药卫生科技项目(No.2021KY947);杭州市医药卫生科技项目(No.B20220028,No.B20220431)。

摘　　要：目的:比较人肺腺癌顺铂耐药株A549/DDP细胞与人肺腺癌顺铂敏感株A549细胞的自噬水平,观察微小RNA-126(microRNA-126,miRNA-126)对肺癌顺铂耐药细胞自噬的影响,并探讨miRNA-126在肺癌顺铂耐药中的作用。方法:MTT法检测顺铂的半数抑制浓度(half maximal inhibitory concentration,IC50);RT-qPCR检测miRNA-126转染后肺癌细胞中miRNA-126的表达量;吖啶橙染色观察肺癌细胞自噬囊泡的形成情况;Western blot检测自噬标志蛋白微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)的表达变化。结果:A549/DDP细胞自噬囊泡数量和LC3水平显著高于A549细胞(P<0.01);转染miRNA-126上调了A549/DDP细胞中miRNA-126的表达量(P<0.01);miRNA-126降低A549/DDP细胞中自噬囊泡比例和LC3-II蛋白表达水平(P<0.01),从而抑制了顺铂耐药细胞A549/DDP的自噬水平;自噬抑制剂3-甲基腺嘌呤(3-methyladenine,3-MA)提高A549/DDP细胞对顺铂的敏感性,miRNA-126联合3-MA进一步增加A549/DDP细胞对顺铂的敏感性(P<0.01)。结论:miRNA-126通过抑制自噬逆转了肺癌A549/DDP细胞顺铂耐药。AIM:Autophagy levels in lung cancer cisplatin-resistant A549/DDP cells and cisplatin-sensitive A549 cells were compared.The effects of microRNA-126(miRNA-126)on autophagy in lung cancer cisplatin-resistant cells were assessed,and the role of miRNA-126 in lung cancer cisplatin-resistant cells was discussed.METHODS:An MTT assay was used to detect the half maximal inhibitory concentration of cisplatin.The miRNA-126 expression was detected by RT-qPCR.Autophagolysosomes were observed under a fluorescence microscope using acridine orange staining.Protein levels of autophagy-associated molecule microtubule-associated protein 1 light chain 3(LC3)in lung cancer cells were determined by Western blot.RESULTS:Autophagy was significantly elevated in A549/DDP cells compared with A549 cells(P<0.01).The expression level of miRNA-126 was significantly elevated in A549/DDP cells after transfection with miRNA-126(P<0.01).miRNA-126 reversed cisplatin resistance of A549/DDP cells,and the level of intracellular acid dye follicular bright red fluorescence in the A549/DDP cells was significantly reduced after transfection with miRNA-126(P<0.01).The LC3-II protein level in A549/DDP cells was significantly reduced after transfection with miRNA-126(P<0.01),and inhibition of cell autophagy by 3-methyladenine(3-MA)enhanced cisplatin sensitivity of A549/DDP cells(P<0.01).Compared with 3-MA+NC group,miRNA-126 combined with 3-MA further increased cisplatin sensitivity of A549/DDP cells(P<0.01).CONCLUSION:miRNA-126 reverses the cisplatin resistance in lung cancer A549/DDP cells by inhibiting autophagy.

关 键 词：微小RNA-126 肺癌 自噬 顺铂 耐药性 

分 类 号：R734.2[医药卫生—肿瘤] R979.11[医药卫生—临床医学] R363