阿瓦斯汀通过miR-664b-3p靶向Beclin1调控非小细胞肺癌细胞A549的自噬水平  

Bevacizumab Regulates Autophagy in Non-small Cell Lung Cancer Cell A549 by Targeting Beclin1 with mir-664b-3p

作　　者：刘劲松[1] 於海军[2] 李丽萍[1] 刘俊[1] 李小菊 LIU Jinsong;YU haijun;LI Liping;LIU Jun;LI Xiaoju(Department of Oncology,First People's Hospital Xiantao of Affiliated to Yangtze University,Xian-tao,Hubei,433000,China;Department of Radiotherapy and Chemotherapy,Zhongnan Hospital of Wuhan University,Wuhan,430061,China)

机构地区：[1]长江大学附属仙桃市第一人民医院肿瘤科,湖北省仙桃市433000 [2]武汉大学中南医院放化疗科,武汉市430061

出　　处：《医学分子生物学杂志》2020年第4期313-317,共5页Journal of Medical Molecular Biology

基　　金：湖北省卫生计生科研项目(No.WJ2018H184)。

摘　　要：目的探讨阿瓦斯汀(Bevacizunab)通过miR-664b-3p靶向Beclinl调控非小细胞肺癌细胞A549的自噬水平的机制。方法通过检测自噬相关基因检测Bevacizumab和其他VEGF抗体对A549细胞的自噬水平的影响;实时荧光定量PCR检测Bevacizumab处理对miR 664b-3p相对表达量的影响;miRDB在线分析miR-664b-3p与Beclin1的相关性;荧光素酶报告系统检测miR-664b-3p是否靶向Beclin1;在过表达或敲低miR-664b-3p的情况下,检测A549细胞的自噬水平。结果使用Bevacizumab处理A549细胞后,自噬相关基因ULK1、BECLIN1、PIK3C3、ATG5、ATG7、LC3B、ATC4B、ATG16L、ATG12的mRNA表达量均显著上升(P均<0.01)。使用Bevacizumab和VEGF抗体处理A549细胞后,其Beclin1和LC3-Ⅱ的表达量上升,自噬水平上升,miR-664b-3p的表达降低。过表达miR-664b-3p后,A549细胞的Beclin1和LC3-Ⅱ的表达量下降,A549细胞自噬水平下降。敲低miR-664b-3p后,A549细胞的Beeclin1和LC3-Ⅱ的表达量上升,自噬水平上升。结论Bevacizumab通过miR-664b-3p靶向Beclin1促进非小细胞肺癌细胞A549的自噬水平。Objective To explore the mechanism of Bevacizumab targeting Beclin1 through miR 664b-3p to regulate the autophagy level of non-small cell lung cancer cell A549.Methods The effects of Bevacizumab and other VEGF antibodies on the autophagy level of A549 cells were examined by detecting autophagy-related genes.Realtime fluorescent quantitative PCR was used to detect the effect of Bevacizumab treatment on the relative expression of miR-664b-3p.miR-664b-3p was analyzed online by miRDB Correlation with Beclin1.Luciferase reporter system was utilized to detect whether miR-664b-3p targets Beclin1.In the case of overexpression or knockdown of miR-664b-3p,the autophagy level of A549 cells was detected.Results After treating A549 cells with Bevacizumab,the mRNA expression of autophagy-related genes ULK1,BECLIN1,PIK3 C3,ATG5,ATG7,LC3 B,ATG4 B,ATG16 L and ATG12 increased significantly(P<0.01 for all).After treating A549 cells with Bevacizumab and VEGF antibodies,the expression of Beclin1 and LC3-Ⅱincreased,the level of autophagy increased,and the expression of miR-664b-3p de-creased.After overexpression of miR.664b-3p,the expression of Beclin1 and LC3-Ⅱin A549 cells decreased,and the autophagy level of A549 cells also decreased.After knocking-down of miR-664b-3p,the expression of Beclin1 and LC3-Ⅱin A549 cells and the level of autophagy were increased.Conclusion Bevacizumab targets Beclin1 through miR-664b-3p to promote the autophagy level of non-small cell lung cancer cells A549.

关 键 词：阿瓦斯汀 miR-664b-3p BECLIN1 非小细胞肺癌 A549 自噬 

分 类 号：R392.11[医药卫生—免疫学]