miR-573调控神经导向因子4促进肺癌细胞上皮间质转化  

iR-573 promotes epithelial-mesenchymal transition of lung cancer cells by regulating Netrin-4

作　　者：肖磊[1] 许丹媛[1] 段林立 何梦璋[1] XIAO Lei;XU Danyuan;DUAN Linli;HE Mengzhang(Department of Respiratory Medicine,Panyu District,the Second Affiliated Hospital of Guangzhou Medical University,Guangdong Guangzhou 510140,China)

机构地区：[1]广州医科大学附属第二医院番禺院区呼吸内科,广东广州510140

出　　处：《现代肿瘤医学》2023年第12期2228-2234,共7页Journal of Modern Oncology

摘　　要：目的:探讨miR-573通过调控神经导向因子4(NTN4)对肺癌细胞上皮间质转化(EMT)的影响。方法:实时荧光定量PCR(qRT-PCR)法检测人肺癌组织和细胞系中miR-573和NTN4 mRNA水平。采用Lipofectamine 2000转染试剂对肺癌A549细胞进行转染并分组为Control组(不转染)、NC inhibitor组(转染NC inhibitor)、miR-573 inhibitor组(转染miR-573 inhibitor)、miR-573 inhibitor+si-NC组(转染miR-573 inhibitor和si-NC)和miR-573 inhibitor+si-NTN4组(转染miR-573 inhibitor和si-NTN4)。qRT-PCR检测miR-573表达水平;划痕法检测迁移能力;Transwell法检测侵袭、迁移能力;蛋白免疫印迹(Western blot)法检测NTN4及E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)表达水平;双荧光素酶实验验证miR-573和NTN4的靶向关系。结果:肺癌组织和细胞系中miR-573表达明显升高,NTN4 mRNA表达明显降低(P<0.05),选择miR-573表达最高的A549细胞进行转染实验。与Control组或NC inhibitor组相比,miR-573 inhibitor组A549细胞中miR-573相对表达量、N-cadherin和Vimentin蛋白水平明显下降,划痕愈合率明显降低,迁移、侵袭细胞数明显减少,E-cadherin蛋白水平显著上升(P<0.05)。miR-573负向调控NTN4表达(P<0.05)。转染si-NTN4可逆转miR-573 inhibitor对A549细胞迁移、侵袭、EMT的抑制作用(P<0.05)。结论:miR-573靶向负调控NTN4表达,促进肺癌细胞的迁移、侵袭和EMT。Objective:To investigate the effect of miR-573 on the epithelial-mesenchymal transition(EMT)of lung cancer cells by regulating Netrin-4(NTN4).Methods:Real-time fluorescent quantitative PCR(qRT-PCR)was used to detect the levels of miR-573 and NTN4 mRNA in human lung cancer tissues and cell lines.Lung cancer A549 cells were transfected with Lipofectamine 2000 transfection reagent and grouped into control group(not transfected),NC inhibitor group(transfected with NC inhibitor),miR-573 inhibitor group(transfected with miR-573 inhibitor),miR-573 inhibitor+si-NC group(transfected with miR-573 inhibitor and si-NC)and miR-573 inhibitor+si-NTN4 group(transfected with miR-573 inhibitor and si-NTN4).qRT-PCR was used to detect the expression level of miR-573.Scratch method was used to detect the migration ability.Transwell cell method was used to detect the invasion and migration abilities.Western blot was used to detect the expression levels of NTN4,E-cadherin,N-cadherin and Vimentin.Dual luciferase experiment was used to verify the targeting relationship between miR-573 and NTN4.Results:The expression of miR-573 was significantly increased and the expression of NTN4 mRNA was significantly decreased in lung cancer tissues and cell lines(P<0.05).The A549 cells with the highest relative expression of miR-573 were selected for transfection experiments.Compared with the control group or NC inhibitor group,the relative expression of miR-573 and the protein levels of N-cadherin and Vimentin in the A549 cells of the miR-573 inhibitor group were significantly decreased.The scratch healing rate was significantly reduced.The numbers of migrating and invading cells were significantly reduced,and the level of E-cadherin protein was significantly increased(P<0.05).miR-573 negatively regulated NTN4 expression(P<0.05).Transfection of si-NTN4 could reverse the inhibitory effects of miR-573 inhibitor on migration,invasion and EMT of A549 cells(P<0.05).Conclusion:miR-573 targets and negatively regulates the expression of NTN4 and promotes th

关 键 词：miR-573 神经导向因子4 肺癌 上皮间质转化 

分 类 号：R734.2[医药卫生—肿瘤]