摘要
Abstract
Objective:To explore effect of miRNA-24 on proliferation and apoptosis of the gastric cancer AGS cell and its potential mechanism.Methods:Expressions of miRNA-24 in gastric cancer AGS,MKN and HGC27 cells as well as gastric mucosal epithelium GES-1 cell were detected by qRT-PCR.The AGS cell line with over-expression of miRNA-24 was constructed.CCK-8,flow cytometry and Western blotting assays were respectively used to measure proliferation vibility of the cells,cell cycle and apoptosis,and expressions of cell cycle and apoptosis-related cyclin D1,CDK2,Bcl-2,p-Iκ B-α/Iκ B-α,p-Rb/Rb proteins.Possible target gene of miRNA-24 was forecasted and verified with dual luciferase report gene assay.Results:Expressions of miRNA-24 in the gastric cells was obviously lower than that in the GES-1 cell (P＜0.05).Proliferation vibility of the AGS cell at 48 h in the transfected with miRNA-24 mimic (miR-24) group was remarkably lower than that in the control miR-NC group ([119.62±12.63]％ vs [147.79±11.89]％,P＜0.05),an arrest of cell cycle occurred,and its early and later apoptosis rates were evidently more increased than those in the miR-NC group (early apoptosis rate:[11.32+2.27]％ vs [0.57±0.08]％;later apoptosis rate:([15.56±2.27]％ vs [0.85±0.16]％,all P＜0.05).In the miRNA-24 group,expression levels of cyclin D1,CDK2,Bcl2 and p-Rb proteins were all obviously lower than those in the miR-NC group,and expression level of pIα.B-α protein was obviously higher than that in the miR-NC group.In co-transfection of miRNA-24 mimic and over-expression plasmid of CARMA3 that may be action target of miRNA-24 (miR-24+pcDNA-CARMA3) group,CARMA3 protein expression of the AGS cell was significantly higher than that in the miR-24 group (1.74±0.09 vs 1.03±0.06,P＜0.05).Proliferation vibility of the AGS cell at 48 h in the miR-24+pcDNA-CARMA3 group was obvi ously higher than that in the miR-24 group ([137.85±15.34]％ vs [102.31±11.23]％,P＜0.05),however,early and later apoptosis rates of the AGS cell in the miR-24+pcDNA-CARMA3 group were all lower than those in the miR-24 group (early apoptosis rate:[4.24±0.56]％ vs [11.32±2.27]％,later apoptosis rate:[6.38+0.63]％ vs [15.56±2.27]％,all P＜0.05).Effect of miRNA-24 on proliferation of apoptosis of the gastric cancer AGS cell can be partially reversed by over-expression of CARMA3.Conclusion:miRNA-24 could inhibit proliferation of the gastric caner cells and promote their apoptosis through targeting CARMA3.关键词
微小RNA-24/胃癌细胞株AGS/增殖/凋亡/半胱天冬酶募集结构域膜相关乌苷酸激酶蛋白3Key words
miRNA-24/gastric cancer/AGS cell/proliferation/apoptosis/CARD recruited membrane associated protein 3 (CARMA3)分类
医药卫生
