Effect Of MiR-181d On Biological Behavior Of Laryngeal Carcinoma Hep2 Cells And Its Molecular Mechanism

Objective:To investigate the effect of miR-181 d on the biological behavior of Hep2 laryngeal cancer cells;To verify whether miR-181 d participates in down-regulating the expression of LCRG1.Methods:1.Potential binding sites for the miR-181 d and 3'UTRs of the LCRG1 gene were predicted using software and further verified by dual-luciferase assay.2.The expression of LCRG1 protein in the mimics/inhibitor NC control group,the miR-181 d mimics/inhibitor experimental group and the blank control group were determined by Western blot.3.The cell proliferation,migration and invasion of miR-181 d mimics/inhibitor Hep2 cells in an experimental group and a blank group were determined by MTT assay,scratch assay,in vitro cell migration assay and in vitro invasion assay.Change in ability.4.Flow cytometry was used to detect the cell cycle.Flow cytometry was used to detect the apoptosis of mimics/inhibitor NC group,miR-181 d mimic/inhibitor group,and blank group.Results:1.Dual luciferase reporter assay results: The first binding sitebetween miR-181 d and LCRG1 3'UTR may be a binding site for miR-181 d.2.The results of Western blot showed that the expression of LCRG1 protein in mimics NC group and blank group had no significant difference(P> 0.05),and the expression of LCRG1 protein in miR-181 d mimics group was significantly lower than that of mimics NC group and blank group(P<0.05).The expression of LCRG1 in miR-181 d inhibitor group was significantly higher than that in inhibitor NC group and blank group(P<0.05).3.MTT assay,scratch assay,in vitro cell migration assay,in vitro invasion assay: Compared with NC control group and blank control group,Hep2 cells transiently transfected with miR-181 d mimic significantly increased migration and invasion of cells,(P<0.05).However,miR-181 d inhibitor transiently decreased the number of migrated and infiltrated cells and significantly inhibited the migration and invasion(P<0.05).4.The results of flow cytometry showed that the percentage of G1 phase and apoptosis rate in miR-181 d inhibitor group were higher than that in inhibitor NC and blank group,and the cell cycle was mainly inhibited in G1 phase.Conclusion:The results suggeste that miR-181 d can bind to LCRG1 3'UTR in Hep2 cells,which may negatively regulate the expression of LCRG1 andenhance the proliferation,migration and invasion of Hep2 cells.