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

发布时间：2018-11-07 15:32

【摘要】：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(P0.05)。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(P0.05)。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(P0.05)。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(P0.05)。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(P0.05)。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。
[Abstract]:Objective to investigate whether upregulation of miR-22 by diallyl disulfide (diallyl disulfide,DADS) inhibits proliferation and migration of human gastric cancer MGC803 cells through Wnt-1 pathway. Methods the effects of DADS and miR-22 on the proliferation and migration and invasion of MGC803 cells were detected by MTT, cell scratch assay and invasion assay respectively. On-line prediction software was used to search for target genes regulated by miR-22, luciferase reporter gene was used to detect the effect of miR-22 on the activity of Wnt-1 3'UTR fluorescence enzyme. QRT-PCR detection of Wnt-1mRNA expression change. Western blot detection of Wnt-1, 尾-catenin and TCF-4 protein expression. Results MTT showed that DADS and miR-22 could significantly inhibit the proliferation of MGC803 cells (P 0.05). Scratch test showed that DADS and miR-22 could significantly inhibit the migration of MGC803 cells, while miR-22 DADS was more obvious (P0.05). Invasion assay showed that miR-22 could inhibit the invasion of MGC803 cells, but miR-22 DADS was more obvious (P0.05). On-line prediction software for target genes regulated by miR-22 showed that Wnt-1 might be the target gene of miR-22. The detection of fluorescein reporter gene confirmed that Wnt-1 was the target gene directly regulated by miR-22. QRT-PCR showed that DADS and miR-22 could down-regulate the expression of Wnt-1 mRNA, but miR-22 DADS was more obvious (P0.05). Western blot showed that DADS and miR-22 could down-regulate the expression of Wnt-1, 尾 -catenin and TCF-4 protein. MiR-22 DADS was especially obvious (P0.05). Conclusion DADS can up-regulate the proliferation and migration and invasion of MGC803 cells via Wnt-1 pathway.
【作者单位】：南华大学肿瘤研究所湖南省胃癌研究中心湖南省高校肿瘤细胞与分子病理学重点实验室;永州职业技术学院基础医学部;南华大学附属湘潭医院病理科;南华大学附属第二医院病理科;
【基金】：国家自然科学基金资助项目(No 81374013,81102854,31100935)
【分类号】：R735.2

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