摘要
目的:研究miR-144-3p对肺癌细胞A549迁移、侵袭及自噬的影响,并探讨其机制。方法:运用qRT-PCR检测细胞中miR-144-3p、EGFR的mRNA表达;将miR-144-3p组(转染miR-144-3p mimics)、miR-NC组(转染miR-control)、si-NC组(转染si-control)、siEGFR组(转染siEGFR)、miR-144-3p+pcDNA 3.1组(miR-144-3p mimics和pcDNA 3.1共转染)、miR-144-3p+EGFR组(miR-144-3p mimics和pcDNA 3.1-EGFR共转染),均用脂质体法转染至A549细胞;Western blot检测各组细胞中EGFR、LC3Ⅱ、LC3Ⅰ、Beclin1的蛋白表达;Transwell检测各组细胞的迁移、侵袭;双荧光素酶报告基因检测实验检测各组细胞的荧光活性。结果:与人正常肺上皮细胞BEAS-2B相比,人肺腺癌细胞A549中miR-144-3p表达显著降低,EGFR表达显著升高(P<0.05);过表达miR-144-3p、敲减EGFR均可显著抑制A549细胞迁移、侵袭,显著升高LC3Ⅱ/LC3Ⅰ、Beclin1的蛋白表达(P<0.05);EGFR是miR-144-3p的靶标。过表达EGFR可逆转miR-144-3p对A549细胞迁移、侵袭的抑制及自噬的促进作用。结论:miR-144-3p可抑制肺癌细胞迁移、侵袭并促进自噬,其机制可能与靶向负调控EGFR有关,将为肺癌的治疗提供新靶点。
Objective:To study the effect of miR-144-3 p on migration,invasion and autophagy of lung cancer cell A549,and explore its mechanism.Methods: qRT-PCR was used to detect the mRNA expression of miR-144-3 p and EGFR in the cells;miR-144-3 p group(transfected miR-144-3 p mimics),miR-NC group(transfected miR-control),si-NC group(trasfeted si-control),siEGFR group(transfected siEGFR),miR-144-3 p+pcDNA 3.1(co-transfected miR-144-3 p mimics and pcDNA 3.1),miR-144-3 p+EGFR group(co-transfected miR-144-3 p mimics and pcDNA 3.1-EGFR) were all transfected into A549 cells by liposome method.Western blot was used to detect the protein expression of EGFR,LC3Ⅱ,LC3Ⅰ and Beclin1 of each group.Transwell was used to detect the migration and invasion of each group.The dual luciferase reporter gene assay was used to detect the fluorescence activity of each group.Results: Compared with human normal lung epithelial cells BEAS-2 B,the expression of miR-144-3 p was significantly decreased in human lung adenocarcinoma cell line A549,and the expression of EGFR was significantly increased(P<0.05).Overexpression miR-144-3 p and knockdown EGFR were all significantly decreased migration and invasion of A549 cells,significantly increased the protein expression of LC3Ⅱ,LC3Ⅰ and Beclin1(P<0.05).EGFR was the target of miR-144-3 p.Overexpression EGFR rescued the inhibition on cell migration and invasion of A549 cells and the promotion on autophagy by miR-144-3 p.Conclusion: miR-144-3 p could inhibit the migration and invasion of lung cancer cells and promote autophagy.The mechanism may be related to the targeted negative regulation of EGFR,which will provide a new target for the treatment of lung cancer.
作者
于林楠
张奎全
祁啸
高爽
任立新
YU Lin-Nan;ZHANG Kui-Quan;QI Xiao;GAO Shuang;REN Li-Xin(Department of General,the Third Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第20期2504-2508,共5页
Chinese Journal of Immunology
作者简介
于林楠,男,医学硕士,主治医师,主要从事肿瘤诊治研究,E-mail:linnanyu1984@163.com;通讯作者:任立新,女,指导教师,主任医师,主要从事肿瘤临床诊治研究,E-mail:renlixin1966@126.com。
