miR-648靶向Syk对胃癌细胞增殖、凋亡、迁移、侵袭的影响及机制研究

Effects of miR-648 targeting Syk on proliferation, apoptosis, migration and invasion of gastric cancer cells and its mechanism

目的研究miR-648在胃癌细胞中的表达及对胃癌细胞增殖、凋亡、迁移和侵袭的影响,并探讨其分子机制。方法采用RT-PCR法和Western blot法检测胃癌细胞和正常胃黏膜上皮细胞中miR-648、脾酪氨酸激酶(spleen tyrosine kinase,Syk)mRNA和Syk蛋白的表达情况。采用Lipofectamine 2000转染胃癌细胞干扰miR-648的表达,采用双荧光素酶基因报告实验检测miR-648对Syk的靶向作用。Western blot法检测转染后胃癌细胞中相关蛋白的表达,CCK-8法、流式细胞术和Transwell法检测细胞活性、凋亡、迁移和侵袭。结果与正常胃黏膜上皮GES-1细胞相比,胃癌SGC-7901、MGC-803细胞中miR-648的表达量显著上调,Syk mRNA和蛋白的表达显著下调。抑制miR-648的表达抑制胃癌SGC-7901细胞的增殖、侵袭、迁移并促进其细胞的凋亡,且Cyclin D1、Bcl-2、MMP-2蛋白的表达量明显下降,Syk、P21、Bax、E-cadherin蛋白的表达量明显上升。miR-648靶向负向调控Syk的表达。沉默Syk可逆转抑制miR-648对胃癌SGC-7901细胞增殖、凋亡、侵袭和迁移的影响。结论miR-648可通过靶向Syk调控胃癌细胞增殖、凋亡、迁移和侵袭。

Objective To study the expression of miR-648 in gastric cancer cells and its effects on proliferation,apoptosis,migration and invasion of gastric cancer cells,and to explore its molecular mechanism.Methods The expressions of miR-648,Syk mRNA and Syk protein in gastric cancer cells and normal gastric mucosal epithelial cells were detected by RT-PCR and western blot.Transfection of gastric cancer cells with Lipofectamine2000 to interfere with the expression of miR-648,and the dual luciferase gene reporter assay was applied to detect the targeting effect of miR-648 on Syk.The expression of related proteins in gastric cancer cells was detected by Western blot.CCK-8,flow cytometry and Transwell were used to detect cell viability,apoptosis,migration and invasion.Results Compared with normal gastric epithelial GES-1 cells,the expression levels of miR-648 were significantly up-regulated in gastric cancer SGC-7901 and MGC-803 cells,and the expression levles of Syk mRNA and protein were significantly down-regulated.Inhibition of miR-648 inhibited the proliferation,invasion,migration and promoted the apoptosis of gastric cancer SGC-7901 cells,and the expression levels of Cyclin D1,Bcl-2 and MMP-2 proteins were decreased significantly,while the expression level of Syk,P21,Bax,E-cadherin proteins were increased significantly.MiR-648 negatively regulates the expression of Syk.Silencing Syk reversed the effects of miR-648 inhibition on proliferation,apoptosis,invasion and migration of gastric cancer SGC-7901 cells.Conclusion miR-648 can regulate the proliferation,apoptosis,migration and invasion of gastric cancer cells by targeting Syk.