Hsa-miR-1827 Inhibits The Migration Of Lung Adenocarcinoma A549 Cell Line And Regulates The Expression Of C-Myc

ObjectiveTo investigate the effect of has-miR-1827 on the migration and proliferative activity of lung adenocarcinoma A549 cell line and whether it regulates the expression of proto-oncogene c-Myc.Methods1.TargetScan7.2 and DIANA-LAB online miRNA target gene prediction software were used to predict the targeted pairing relationship between hsa-miR-1827 and c-Myc mRNA 3'untranslated region(UTR).TargetScan7.2,DIANA-LAB,and miRDB were used to predict the target genes of hsa-miR-1827 respectively and then their intersection was acquired by venny2.1,followed by KEGG pathway analysis.The association between hsa-miR-1827 expression and prognosis,pathological stage of lung adenocarcinoma patients was explored using published data from the GEO database.2.The hsa-miR-1827 mimics was transfected into the lung adenocarcinoma A549 cell line using liposomes.Real-Time qPCR was performed to detect the expression of hsa-miR-1827 and c-Myc mRNA after transfection.The change of c-Myc protein expression was detected by Western Blot.We explored the effect of hsa-miR-1827 on the migration function of A549 using Transwell migration assay and cell scratch assay,we also conducted cell counting kit-8 assay to explore the effect of hsa-miR-1827 on the proliferative ability of A549.A549 cells transfected with the miR-NC sequence were used as the control group.3.The dual luciferase reporter gene assay was used to identify whether hsa-miR-1827 directly binds to c-Myc mRNA 3'UTR and confirm the specific binding sites.Results1.TargetScan7.2 and DIANA-LAB showed that there is a mutual pairing sequence between the hsa-miR-1827 seed region and the c-Myc mRNA 3'UTR region.After target gene prediction and KEGG pathway analysis,it is suggested that some of target genes of has-miR-1827 are involved directly or indirectly in tumor progression.The data from GEO chip GSE63805 showed that the 10-year overall survival rate(after surgery)of patients diagnosed as lung adenocarcinoma with higher expression of hsa-miR-1827 was significantly better than that of patients with lower expression of hsa-miR-1827(37.5%vs.6.25%).2.Compared with the miR-NC transfected group,the expression of hsa-miR-1827 was significantly up-regulated,and the mRNA and protein levels of c-Myc were significantly decreased in the hsa-miR-1827 transfected group.hsa-miR-1827 could significantly inhibit the migration function of A549,but not proliferative ability.3.The dual luciferase reporter assay suggested that hsa-miR-1827 downregulated c-Myc expression by pairing with c-Myc mRNA 3' UTR(139-146 bases)directly.Conclusionhsa-miR-1827 can regulate the expression of proto-oncogene c-Myc in lung adenocarcinoma and inhibit the migration of A549 cell line.