miRNA-299-3p对胃癌AGS细胞增殖、凋亡、迁移的影响及其机制  

Effect and Mechanism of miRNA-299-3p on the Proliferation,Apoptosis,and Migration of Gastric Cancer AGS Cells

作　　者：范金阳 杜波涛 程志通 FAN Jinyang;DU Botao;CHENG Zhitong(Department of Surgical Oncology,Xinxiang Central Hospital,Xinxiang 453000,China)

机构地区：[1]新乡市中心医院肿瘤外科,河南新乡453000

出　　处：《河南医学研究》2025年第2期198-203,共6页Henan Medical Research

摘　　要：目的探索miRNA-299-3p对胃癌AGS细胞增殖、凋亡、细胞周期及迁移的影响,并进一步探索其机制。方法体外培养胃癌AGS细胞,并将其分为空白对照组、阴性对照组及实验组。利用qPCR检测miRNA-299-3p的表达,将空载体质粒和miRNA-299-3p质粒分别转染入AGS细胞;利用CCK-8法检测细胞增殖情况;流式细胞术检测细胞凋亡、细胞周期及活性氧水平变化情况;Transwell法检测细胞迁移和侵袭情况;Western blot检测相关蛋白表达量变化情况。结果CCK-8实验表明,miRNA-299-3p的高表达抑制AGS细胞增殖(P<0.05);流式细胞术及Western blot结果显示,miRNA-299-3p的高表达能够诱导AGS细胞凋亡,caspase-3和Bax表达量升高,Bcl-2蛋白表达量降低(P<0.05);同时,miRNA-299-3p过表达后,能够下调CDK1/2和Cyclin B1蛋白的表达量,将AGS细胞周期阻滞在G_(2)/M期;Transwell小室及Western blot实验结果表明,miRNA-299-3p过表达使N-cadherin蛋白表达量降低,E-cadherin蛋白表达量升高从而抑制AGS细胞迁移;另外,miRNA-299-3p过表达能够上调AGS细胞中活性氧水平。结论过表达miRNA-299-3能够通过上调细胞内活性氧水平,抑制AGS细胞的增殖、诱导AGS细胞发生凋亡、将细胞周期阻滞在G_(2)/M期、抑制迁移,为miRNA-299-3p在胃癌研究中提供一定的理论基础。Objective To explore the effect of miRNA-299-3p on proliferation,apoptosis,cell cycle and migration of gastric cancer AGS cells,and to further explore its mechanism.Methods Gastric cancer AGS cells in vitro were culture and divided into blank control group,negative control group and experimental group.The expression of miRNA-299-3p were detected by qPCR,and transfected the empty vector plasmid and miRNA-299-3p plasmid into AGS cells.The CCK-8 was used to detect cell proliferation,and used flow cytometry to detect apoptosis,cell cycle and reactive oxygen levels.The Transwell was used to detect cell migration and invasion.The Western blot was used to detect changes in related protein expression.Results The CCK-8 experiment showed that high expression of miRNA-299-3p inhibited AGS cell proliferation(P<0.05).Flow cytometry and Western blot results showed that high expression of miRNA-299-3p can induce AGS cell apoptosis,caspase-3 and Bax expression increased,Bcl-2 protein expression decreased(P<0.05).Meanwhile,after overexpression of miRNA-299-3p,it was able to down-regulate the expression of CDK1/2 and Cyclin B1 protein to block AGS cell cycle in G_(2)/M phase.Transwell chambers and Western blot experiments showed that overexpression of miRNA-299-3p reduced the expression of N-cadherin protein and increased the expression of E-cadherin protein to inhibit the migration of AGS cells.In addition,the miRNA-299-3p overexpression could be up-regulated reactive oxygen levels in AGS cells.Conclusion Overexpression of miRNA-299-3 can inhibit the proliferation of AGS cells,induce apoptosis of AGS cells,block the cell cycle at G_(2)/M phase,and inhibit migration by up-regulating the level of intracellular reactive oxygen species,and this can provide a certain theoretical basis in gastric cancer research for miRNA-299-3p.

关 键 词：miRNA-299-3p 胃癌AGS细胞 细胞凋亡 细胞周期 活性氧 细胞迁移 

分 类 号：R73[医药卫生—肿瘤]