【摘要】：目的探討二烯丙基二硫(diallyl disulfide,DADS)上調(diào)miR-22是否通過Wnt-1通路抑制人胃癌MGC803細(xì)胞增殖與遷移侵襲。方法 MTT、細(xì)胞劃痕實(shí)驗(yàn)、侵襲實(shí)驗(yàn)分別檢測(cè)DADS與miR-22對(duì)MGC803細(xì)胞增殖與遷移侵襲的影響。在線預(yù)測(cè)軟件尋找miR-22調(diào)控的靶基因,熒光素酶報(bào)告基因檢測(cè)miR-22對(duì)Wnt-1 3'UTR熒光酶活性的影響。qRT-PCR檢測(cè)Wnt-1mRNA表達(dá)變化。Western blot檢測(cè)Wnt-1、β-catenin與TCF-4蛋白表達(dá)。結(jié)果MTT顯示,DADS與miR-22可明顯抑制MGC803細(xì)胞增殖(P0.05)。劃痕實(shí)驗(yàn)顯示,DADS與miR-22可明顯抑制MGC803細(xì)胞遷移,而miR-22+DADS更為明顯(P0.05)。侵襲實(shí)驗(yàn)顯示,miR-22可抑制人胃癌MGC803細(xì)胞侵襲,而miR-22+DADS更為明顯(P0.05)。在線預(yù)測(cè)軟件尋找miR-22調(diào)控的靶基因顯示,Wnt-1可能是miR-22的靶基因,熒光素報(bào)告基因檢測(cè)證實(shí)Wnt-1是miR-22直接調(diào)控的靶基因;qRT-PCR顯示,DADS與miR-22能下調(diào)Wnt-1 mRNA表達(dá),而miR-22+DADS更為明顯(P0.05)。Western blot顯示,DADS與miR-22能下調(diào)Wnt-1、β-catenin與TCF-4蛋白表達(dá),而miR-22+DADS尤為明顯(P0.05)。結(jié)論 DADS可上調(diào)miR-22通過Wnt-1通路明顯抑制MGC803細(xì)胞增殖與遷移侵襲。
[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.
【作者單位】： 南華大學(xué)腫瘤研究所湖南省胃癌研究中心湖南省高校腫瘤細(xì)胞與分子病理學(xué)重點(diǎn)實(shí)驗(yàn)室;永州職業(yè)技術(shù)學(xué)院基礎(chǔ)醫(yī)學(xué)部;南華大學(xué)附屬湘潭醫(yī)院病理科;南華大學(xué)附屬第二醫(yī)院病理科;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(No 81374013,81102854,31100935)
【分類號(hào)】：R735.2

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