Effects Of MiR-141-3p On Growth,Proliferation,Migration And Invasion Of Hep-2 Cells

Objective Laryngeal cancer is a common tumor in the head and neck.Its morbidity and mortality are increasing year by year.Therefore,early diagnosis and treatment of laryngeal cancer is the key to improve the 5 year survival rate of laryngeal cancer patients.In recent years,studies have found that miR-141-3p plays an important role in the proliferation,migration and invasion of gastric cancer,colon cancer and esophageal cancer cells.Thus,we investigated whether mi R-141-3p affects the proliferation,migration and invasion of Hep-2 cells of laryngeal carcinoma and analyzed its possible mechanism.Methods 1.The human LSCC cell line Hep-2 was used as the research object,they were divided into three groups: miR-141-3p mimics group(the Hep-2 cells were transfected with mi R-141-3p mimics),miR-NC group(the Hep-2 cells were transfected negative control RNA)and blank control group(non-transfected).2.Transfected of miR-141-3p mimics or negative control RNA into the Hep-2 cells using Lipofectamine 2000 in accordance with the manufacturer procedure.Cells were transfected by the FAM-labeled miR-NC.After cultured for 6 h,the cells were visualized by fluorescence microscope to evaluate the transfection efficiency.Real-time fluorescence quantitative reaction(RT-PCR)method was used to detect the miR-141-3p transcriptional expression in Hep-2 cells.3.The effects of miR-141-3p on the proliferation,migration and invasion of the cells were also detected using MTT,scratch and Transwell invasion assays,respectively.4.The prediction of ZEB1 mRNA as a target of miR-141-3p was made with Pictar and TargetScan programs,then RT-PCR and Western Blot were analyzed the expression of ZEB1 from mRNA and protein levels respectively.Results 1.After transfection,the expression level of miR-141-3p in the miR-141-3p mimics Hep-2 cells group was significantly increased,miR-141-3p can be expressed in Hep-2 cells of laryngeal carcinoma.2.An MTT assay demonstrated that overexpression of miR-141-3p significantly suppressed the proliferation of Hep-2 cells at 24,48,and 72 h after transfection,respectively(P < 0.05).3.Cells migration in miR-141-3p mimics group was less than those in other group(P < 0.05)by scratch test.4.After transfection at the same time,Fewer Hep-2 cells transfected of miR-141-3p mimics migrated through the porous transwells compared with the negative control cells and the blank control cells.5.Based on the bioinformatics analysis using two different programs(Pictar and targetscan)suggested that ZEB1 mRNA is a potential target of miR-141-3p.6.Western blot and real-time RT-PCR results indicated that both ZEB1 mRNA and protein levels in miR-141-3p mimics group were significantly lower than those in other control groups(P < 0.05).Conclusion 1.The results suggest that the miR-141-3p inhibited the proliferation,migration and invasion of human LSCC Hep-2 cells.2.MiR-141-3p may affect the development of laryngeal carcinoma by downregulating the expression of ZEB1 in Hep-2 cells.