The Effect Of MiR-375 On The Biological Behavior Of Laryngeal Carcinoma Cells And Its Mechanism

Objective: Laryngeal carcinoma is one of the common malignant tumors in the head and neck. In recent years, the incidence of laryngeal cancer is younger and higher. In this study, the human laryngeal cancer cell line as the research object, analysis the effects of miR-375 on the biological behavior of Hep-2 cells, and discuss the possible molecular mechanisms.Methods: This study was divided into miR-375 mimics group(transfected with miR-375), miR-NC group(transfected with nonsense sequence miR-NC) and Blank control group(transfected with nothing).Using Lipofectamine TM 2000 transfected miR-375 and MiR-NC into Laryngeal carcinoma Hep-2 cells by GFP fluorescence to detect the transfection efficiency;MTT assay was used to study cell proliferation;Flow cytometry was used to detecte Cell cycle and apoptosis;Cell scratch test was used to observe Cell migration; Bioinformatics software was used to predict the target genes of miR-375; RT-PCR method and Western Blot method were separately used to detect the expression level of IGF1 R gene mRNA and protein in cells.Results:Inserting miR-375 into Hep-2 cells, the fluorescence detection showed that miR-375 can be highly expressed in Hep-2 cells of miR-375 mimics group; MTT assay detection showed that the Hep-2 cells proliferation rate of miR-375 mimics group was lower than miR-NC group and Blank control group; flow cytometry detection showed thatThe Hep-2 cells sub-G0/G1 phase ratio of miR-375 mimics group was higher than miR-NC group and Blank control group; The Hep-2 cells apoptotic rate of miR-375 mimics group was significantly higher than miR-NC group and Blank control group; the difference was statistically significant(P<0.01); scratch test results showed that The Hep-2 cells relative migration distance of miR-375 mimics group was lower than miR-NC group and Blank control group,the difference was statistically significant(P<0.01); MiRNA target prediction software predicted that IGF1 R may be one of the target genes of miR-375; RT-PCR and Western Blot result showed that the Hep-2 cells relative expression of IGF1 R gene mRNA and protein in miR-375 mimics group was significantly decreased compared with miR-NC group and Blank control group, the difference was statistically significant(P<0.01).Conclusion: 1.Inserting miR-375 into Hep-2 cells, slow the proliferation of Hep-2 cells, induce cell block in the G0/G1 phase,increase cell apoptosis, inhibit migration.2.Inserting miR-375 into Hep-2 cells, the Hep-2 cells relative expression of IGF1 R gene was reduced.