LncRNA DDX11-AS1通过靶向miR-497-5p调控胃癌细胞增殖、迁移和凋亡的机制研究

Mechanism of LncRNA DDX11-AS1 in Regulating Proliferation,Migration and Apoptosis of Gastric Cancer Cells by Targeting MiR-497-5p

背景:LncRNA在胃癌中表达上调或下调,其在胃癌发生、发展中的作用机制尚未完全阐明。目的:探讨LncRNA DDX11-AS1通过靶向miR-497-5p表达来调控胃癌细胞增殖、迁移、凋亡的机制。方法:采用qRT-PCR检测胃癌组织和细胞株中DDX11-AS1和miR-497-5p的表达,Pearson法分析胃癌组织中DDX11-AS1与miR-497-5p的相关性。将胃癌HGC-27细胞随机分为NC组、si-con组、si-DDX11-AS1组、miR-con组、miR-497-5p组、si-DDX11-AS1+anti-miR-con组、si-DDX11-AS1+anti-miR-497-5p组。MTT法检测细胞增殖情况;流式细胞术检测细胞凋亡情况;Transwell实验检测细胞迁移和侵袭能力;双萤光素酶报告系统验证DDX11-AS1与miR-497-5p的靶向调节关系;蛋白质印迹法检测cyclin D1、cleaved caspase-3、MMP-2、MMP-9蛋白表达。结果:胃癌组织和细胞株中DDX11-AS1表达显著升高(P<0.05),而miR-497-5p表达显著降低(P<0.05),DDX11-AS1与miR-497-5p呈负相关(r=-0.754,P<0.05)。干扰DDX11-AS1表达或上调miR-497-5p表达可显著抑制胃癌细胞增殖、迁移和侵袭(P<0.05),诱导细胞凋亡(P<0.05),cyclin D1、MMP-2、MMP-9表达显著降低(P<0.05),cleaved caspase-3表达显著升高(P<0.05)。双萤光素酶报告实验证明DDX11-AS1可负向调控miR-497-5p的表达和活性。抑制miR-497-5p表达可逆转干扰DDX11-AS1表达对胃癌细胞生物学行为的影响。结论:干扰LncRNA DDX11-AS1表达能通过靶向结合并上调miR-497-5p的表达而抑制胃癌细胞增殖、迁移和侵袭,并诱导细胞凋亡。

Background:LncRNA expression was up-regulated or down-regulated in gastric cancer,but the mechanism of role of LncRNA in the development and progression of gastric cancer was not been fully clarified.Aims:To investigate the mechanism of LncRNA DDX11-AS1 in regulating the proliferation,migration and apoptosis of gastric cancer cells by targeting the expression of miR-497-5p.Methods:qRT-PCR was used to detect the expressions of DDX11-AS1 and miR-497-5p in gastric cancer tissue and cell lines.Pearson method was used to analyze the correlation between DDX11-AS1 and miR-497-5p in gastric cancer tissue.Gastric cancer HGC-27 cells were randomly divided into NC group,si-con group,si-DDX11-AS1 group,miR-con group,miR-497-5p group,si-DDX11-AS1+anti-miR-con group,si-DDX11-AS1+anti-miR-497-5p group.The cell proliferation was detected by MTT.The cell apoptosis was detected by flow cytometry.Transwell assay was used to detect the migration and invasion ability.The dual luciferase reporter system assay was used to verify the targeted regulatory relationship between DDX11-AS1 and miR-497-5p.The protein expressions of cyclin D1,cleaved caspase-3,MMP-2 and MMP-9 were detected by Western blotting.Results:The expression of DDX11-AS1 in gastric cancer tissue and cell lines was significantly increased(P<0.05),while the expression of miR-497-5p was significantly decreased(P<0.05),DDX11-AS1 was negatively correlated with miR-497-5p(r=-0.754,P<0.05).Interfering the expression of DDX11-AS1 or up-regulating the expression of miR-497-5p significantly inhibited cell proliferation,migration and invasion(P<0.05),apoptosis rate was significantly increased(P<0.05),and the expressions of cyclin D1,MMP-2 and MMP-9 were significantly decreased(P<0.05),and the expression of cleaved caspase-3 was significantly increased(P<0.05).The dual luciferase reporter assay demonstrated that DDX11-AS1 negatively regulated the expression and activity of miR-497-5p.Inhibition of miR-497-5p expression reversed the effect of interference with DDX11-AS1 expression on the biological behavior of gastric cancer cells.Conclusions:Interference with LncRNA DDX11-AS1 expression can inhibit the proliferation,migration,invasion and induce apoptosis of gastric cancer cells by targeting and up-regulating the expression of miR-497-5p.