| BackgroundLung cancer is a common primary malignant tumor,which has become a major cause of threat to human health.Micro RNA(miRNA)complements the target gene m RNA 3’Untranslated Regions(UTR)in transcription.Post-level negative regulation of m RNA expression was closely related to the occurrence and development of many diseases including lung cancer.Because the miR-203 gene was derived from epithelial basal cells,the study of its correlation with malignant tumors has become the focus.miR-203a was a member of the miR-203 family and had been confirmed to be abnormally low in lung cancer tissues and cells.However,its role in regulating tumor cell metastasis and its molecular mechanism were not completely clear.ObjectiveTo explore the effect of miR-203a on the migration and invasion of non-small cell lung cancer A549 cells and its mechanism,in order to provide new clues and basis for lung cancer target gene therapy.MethodsA549 cells cultured in vitro were divided into control group(untransfected),miR-NC group(transfected miR-203a mimics control),miR-203a group(transfected miR-203a mimics control),anti-miR-NC group(transfected miR-203a inhibitor control)and anti-miR-203a group(transfected miR-203a inhibitor control).The expression of miR-203a in each group was detected by RT-PCR,and cell migration ability was detected by scratch test.The invasive ability of cells was detected by transwell lab test,and the expression of EMT-related proteins E-cadherin and Vimentin was detected by Western blot.Bioinformatics software was used to predict the potential target genes of miR-203a,and dual luciferase reporter gene experiment was used to detect the targeting relationship between miR-203a and Slug,Western blot and RT-PCR were used to detect the regulatory effect of miR-203a on Slug expression.Liposome was used to transfect pc DNA3.1-Slug overexpression plasmid and interference sequence si Slug and its corresponding control pc DNA3.1 and si NC.Scratch experiment,Transwell chamber experiment and Western blot were used to detect the effect of Slug on A549 cells under the control of miR-203a.Migration,invasion and epithelial-mesenchymal transition(EMT)related protein Slug,E-cadherin,Vimentin expression.Results 1.Compared with the control group,the miR-203a group has significantly reduced migration distance(P<0.05),and the number of invaded cells was significantly reduced(P<0.05),while the anti-miR-203a group was significantly higher than the control group(P<0.05).2.Compared with the control group,the expression level of E-cadherin protein in the miR-203a group was significantly increased,while the expression level of Vimentin protein was significantly reduced(P<0.05).Compared with the control group,anti-miRThe expression level of E-cadherin protein in the 203a group was significantly reduced,while the expression level of Vimentin protein was significantly increased(P<0.05).3.The bioinformatics software predicts that Slug 3’UTR had a binding site that can complement miR-203a.The dual luciferase reporter gene confirms that Slug was a potential target gene of miR-203a.4.Compared with the control group,the expression of Slug protein and m RNA in the miR-203a group was significantly reduced(P<0.05),while the expression of Slug protein and m RNA in the anti-miR-203a group was significantly increased(P<0.05).5.Compared with the control group,the miR-203a group’s migration distance,the number of invasive cells,and the expression of Slug and Vimentin proteins were significantly reduced,while the expression of E-cadherin protein was significantly increased(P<0.05).Compared with the miR-203a group Compared with the miR-203a+pc DNA group,there was no significant change(P>0.05),but in the miR-203a+Slug group,the migration distance,the number of invaded cells,and the expression of Slug and Vimentin proteins were significantly increased,while the expression of E-cadherin protein was significantly decreased(P<0.05).6.Compared with the control group,in the anti-miR-203a group,the migration distance,the number of invasive cells,and the expression of Slug and Vimentin proteins were significantly increased,while the expression of E-cadherin protein was significantly decreased(P<0.05),there was no significant difference between the anti-miR-NC group and the control group(P>0.05).Compared with the anti-miR-203a group,the anti-miR-203a+si NC group had no significant change(P>0.05),but the anti-miR-203a In the +si Slug group,the migration distance,the number of invasive cells,and the expression of Slug and Vimentin proteins were significantly reduced,while the expression of E-cadherin protein was significantly increased(P<0.05).Conclusion1.Mir-203a could inhibit migration and invasion of A549 cells.2.The mechanism of inhibition of migration and invasion of A549 cells by miR-203a may be related to the inhibition of EMT process.3.Slug was a potential target gene of miR-203a.4.miR-203a could inhibit the migration and invasion of lung cancer A549 cells by targeting Slug to regulate EMT. |
