miR-573调控神经导向因子4促进肺癌细胞上皮间质转化

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...

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

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 the migration,invasion and EMT of lung cancer cells.