摘要
目的探究长链非编码RNA(lncRNA)ANCR调控miR-497-5p对胃癌细胞血管管腔形成、凋亡及PI3K/AKT信号通路的影响。方法取MGC803胃癌细胞将其分为胃癌细胞(GC)组、胃癌细胞+lncRNA ANCR-NC(AN)组、胃癌细胞+lncRNA ANCR过表达质粒(AM)组、胃癌细胞+lncRNA ANCR siRNA(AI)组、胃癌细胞+miR-497-5p-NC(MN)组、胃癌细胞+miR-497-5p siRNA(MI)组、胃癌细胞+miR-497-5p mimics(MM)组、胃癌细胞+lncRNA ANCR siRNA+miR-497-5p mimics(IM)组,管腔形成实验观察细胞血管管腔形成,PI染色法检测细胞凋亡,免疫印迹法检测PI3K和AKT蛋白表达,双荧光素酶报告实验验证lncRNA ANCR和miR-497-5p的相互作用。结果与AN组、MN组相比,AM组、MI组凋亡率明显降低(P<0.05),血管管腔形成数量、PI3K和AKT蛋白表达明显升高(P<0.05);与AM组、MI组相比,AI组、MM组凋亡率明显升高(P<0.05),血管管腔形成数量、PI3K和AKT蛋白表达明显降低(P<0.05);与AI组、MM组相比,IM组凋亡率明显升高(P<0.05),血管管腔形成数量、PI3K和AKT蛋白表达明显降低(P<0.05)。双荧光素酶报告结果显示,转染lncRNA ANCR后可显著降低miR-497-5p-3′-UTR-WT的荧光素酶活性(P<0.05),但对突变基因无影响(P>0.05)。结论抑制lncRNA ANCR可有效抑制胃癌细胞血管管腔形成,促进细胞凋亡,并抑制PI3K和AKT信号通路表达,其作用机制可能与靶向激活miR-497-5p有关。
Objective To explore the effects of lncRNA ANCR regulating miR-497-5p on vascular lumen formation,apoptosis and PI3K/AKT signaling pathway of gastric cancer cells.Methods Gastric cancer cells were collected and divided into gastric cancer cell(GC)group,gastric cancer cell+lncRNA ANCR-NC(AN)group,gastric cancer cell+lncRNA ANCR overexpressed plasmid(AM)group,gastric cancer cell+lncRNA ANCR siRNA(AI)group,gastric cancer cells+miR-497-5p-NC(MN)group,gastric cancer cells+miR-497-5p siRNA(MI)group,gastric cancer cells+miR-497-5p mimics(MM)group,and gastric cancer cells+lncRNA ANCR siRNA+miR-497-5p mimics(IM)group.Vascular lumen formation was observed by lumen formation assay,cell apoptosis was detected by PI staining method,and protein expression of PI3K and AKT was detected by western blotting.The interaction between lncRNA ANCR and miR-497-5p was confirmed by double luciferase reporting assay.Results Compared with AN group and MN group,the apoptosis rate of AM group and MI group was significantly decreased(P<0.05),the number of vascular lumen formation and the expression of PI3K and AKT protein were significantly increased(P<0.05).Compared with AM and MI groups,the apoptosis rate of AI and MM groups was significantly increased(P<0.05),while the number of vascular lumen formation and the expression of PI3K and AKT protein were significantly decreased(P<0.05).Compared with AI and MM groups,the apoptosis rate of IM group was significantly increased(P<0.05),the number of vascular lumen formation and the expression of PI3K and AKT protein were significantly decreased(P<0.05).The results of double luciferase report showed that transfection of lncRNA ANCR could significantly reduce the luciferase activity of miR-497-5p-3′-UTR-WT(P<0.05),but had no significant effect on mutant genes(P>0.05).Conclusion Inhibition of lncRNA ANCR can effectively inhibit the formation of vascular lumen,promote cell apoptosis,and inhibit the expression of PI3K/AKT signaling pathway in gastric cancer cells,which may be related to the targeted activation of miR-497-5p.
作者
朱婷婷
夏一函
孙莎莎
高鹏
邱文生
ZHU Tingting;XIA Yihan;SUN Shasha;GAO Peng;QIU Wensheng(Department of Gastroenterology,Qingdao Eighth People s Hospital,Qingdao,Shandong 266000,China)
出处
《中国临床研究》
CAS
2024年第2期216-222,共7页
Chinese Journal of Clinical Research
基金
国家自然科学基金(81472338)
山东省自然科学基金资助创新发展基金项目(ZR2021LZL001)。
