二烯丙基二硫上调miR-22通过Wnt-1通路抑制人胃癌细胞增殖与迁移侵袭

Up-regulation of miR-22 through Wnt pathway suppresses proliferation,migration and invasion in human gastric MGC803 cells by DADS

目的探讨二烯丙基二硫(diallyl disulfide,DADS)上调miR-22是否通过Wnt-1通路抑制人胃癌MGC803细胞增殖与迁移侵袭。方法 MTT、细胞划痕实验、侵袭实验分别检测DADS与miR-22对MGC803细胞增殖与迁移侵袭的影响。在线预测软件寻找miR-22调控的靶基因,荧光素酶报告基因检测miR-22对Wnt-1 3'UTR荧光酶活性的影响。qRT-PCR检测Wnt-1mRNA表达变化。Western blot检测Wnt-1、β-catenin与TCF-4蛋白表达。结果MTT显示,DADS与miR-22可明显抑制MGC803细胞增殖(P<0.05)。划痕实验显示,DADS与miR-22可明显抑制MGC803细胞迁移,而miR-22+DADS更为明显(P<0.05)。侵袭实验显示,miR-22可抑制人胃癌MGC803细胞侵袭,而miR-22+DADS更为明显(P<0.05)。在线预测软件寻找miR-22调控的靶基因显示,Wnt-1可能是miR-22的靶基因,荧光素报告基因检测证实Wnt-1是miR-22直接调控的靶基因;qRT-PCR显示,DADS与miR-22能下调Wnt-1 mRNA表达,而miR-22+DADS更为明显(P<0.05)。Western blot显示,DADS与miR-22能下调Wnt-1、β-catenin与TCF-4蛋白表达,而miR-22+DADS尤为明显(P<0.05)。结论 DADS可上调miR-22通过Wnt-1通路明显抑制MGC803细胞增殖与迁移侵袭。

Aim To investigate the up-regulation of miR-22 through Wnt pathway inhibits the proliferation,migration and invasion in human gastric MGC803 cells induced by diallyl disulfide（ DADS）. Methods The effects of proliferation,migration,and invasion of gastric cancer cells were evaluated by MTT,wound-healing and invasion assays. Online prediction software was applied to search the target gene of miR-22. Luciferasereport gene assay was used to assess the target genes Wnt-1 of miR-22. The expressions of Wnt-1,β-catenin and TCF-4 were tested by qRT-PCR and Western blot,respectively. Results MTT showed that DADS and miR-22 notably decreased the proliferation compared with control group（ P〈0. 05）. Wound-healing assay showed that DADS and miR-22 could significantly inhibit the migration of MGC 8 0 3 cells compared with thecontrol group, especially in miR-22 ＋ DADS（ P〈0. 05）. Invasion assay showed that DADS and miR-22 could markedly inhibit the invasion of MGC803 cells compared with the control group,especially in miR-22＋ DADS（ P〈0. 05）. Online prediction software to search the target gene exhibited that Wnt-1 may be a target gene of miR-22. Luciferase report gene assay disclosed that Wnt-1 was identified as a direct target of miR-22. qRT-PCR showed that the expression of Wnt-1 mRNA was respectively down-regulated by DADSand miR-22 compared with control group,especially in miR-22 ＋ DADS（ P〈0. 05）. Western blot exhibited that DADS and miR-22 obviously suppressed the expressions of Wnt-1,β-catenin and TCF-4 proteins,especially in miR-22 ＋ DADS（ P〈0. 05）. Conclusion Up-regulation of miR-22 through Wnt pathway can remarkably suppress the proliferation,migration and invasion in MGC803 cells by DADS.