The Expression Level,role And Its Machanism Of MicroRNA-205-5p In Squamous Cell Carcinoma SCC12 Cells

Objective:The aim of this study was to investigate the effect of miR-205-5p on the proliferation of cutaneous squamous cell carcinoma cell line SCC12 in vitro and its association with Bcl-2.Methods:1.The expression of different mi RNA levels in HaCaT cells and SCC12 cells was detected by miRNA microarray Assay.2.Real-time PCR was performed on SCC12 cellsand HaCaT cells to determine miR-205-5p expression;3.The SCC12 cells were transfected with miR-205-5p mimics,miR-205-5p inhibitor,miRNA NC(carrying by Lipofectamine 2000),but no transfection was performed in normal cells.4.Real-time PCR was used to analyze the expression of mi R-205-5p in different cell groups and evaluate the transfection effect about 48 hours.5.CCK8 assay was used to detect the proliferation of SCC12 cells in four groups after 48 hours of transfection.6.Apoptosis of SCC12 cells in four groups was detected by flow cytometry after 48 hours of transfection.7.RT-PCR was used to detect the mRNA levels of Bcl-2 in the cells of each group after 48 hours of transfection.8.After 72 hours of transfection,Western-blotting was used to detect the protein expression of Bcl-2 and Bax.Results:1.RT-PCR showed that miR-205-5p expression in HaCaT cells significantly increased to about 20 fold of that in SCC12 cells(P<0.01).2.After transfection,the expression of mi R-205-5p in miR-205-5p group was significantly higher than that in miR-205-5p inhibitors group,NC group and control group(P < 0.05).The expression of miR-205-5p in miR-205-5p inhibitors group was significantly lower than that in negative control(NC)group,control group and miR-205-5p group(P < 0.05).3.The proliferation of scc12 cells was decreased after tansfected with the miR-205-5p mimics(P<0.05),increased after transfected with the miR-205-5p inhibitors(P < 0.05).4.The apoptosis rate of the SCC12 cells was higher after tansfected with the miR-205-5p mimics(P<0.05),lower after transfected with the miR-205-5p inhibitors(P < 0.05).7.The expression of Bcl-2 at mRNA levels in miR-205-5p group was significantly down-regulated compared with miR-205-5p inhibitors group,NC group and control group(P<0.05).8.The expression of Bcl-2 at protein levels in miR-205-5p group was significantly down-regulated compared with miR-205-5p inhibitors group,NC group and control group(P<0.05).Conclusion:1.miR-205-5p is downregulated in SCC12 cells compared to HaCaT cells.2.miR-205-5p can inhibite Bcl-2 mRNA and protein expression.3.miR-205-5p can significantly inhibit proliferation and induce apoptosis of SCC12 cells,and its mechanism may be through targeting Bcl-2.4.miR-205-5p may serve as a new target to aid in the diagonosis and treatment of SCC.