MicroRNA-146a通过靶向TAK1诱导胃癌SGC-7901细胞凋亡

发布时间：2018-03-28 02:18

本文选题：胃癌SGC-细胞　切入点：微小RNA-a　出处：《中国病理生理杂志》2017年08期

【摘要】：目的:观察微小RNA-146a(miR-146a)对胃癌SGC-7901细胞凋亡的影响及其可能的分子机制。方法:将miR-146a mimic(上调miR-146a表达)和miR-146a inhibitor(下调miR-146a表达)分别转染至人胃癌SGC-7901细胞中,同时设置miRNA无义序列转染为阴性对照组(NC组)。RT-q PCR检测转染后胃癌细胞miR-146a的表达水平;CCK-8法和流式细胞术检测miR-146a对SGC-7901胃癌细胞生长活力和凋亡的影响;RT-q PCR和Western blot法检测miR-146a上调或下调对转化生长因子β激活激酶1(TAK1)/核因子κB(NF-κB)通路的影响。结果:在SGC-7901细胞中,上调miR-146a表达显著促进细胞凋亡,而下调miR-146a表达则显著抑制细胞凋亡。与NC组相比,miR-146a mimics转染SGC-7901细胞后,TAK1的mRNA和蛋白表达均明显下降,差异均有统计学显著性(P0.05),而miR-146a inhibitors转染组TAK1的mRNA和蛋白质表达则显著增加(P0.05),提示miR-146a负调控TAK1表达。敲低TAK1促进SGC-7901细胞凋亡(P0.01),而TAK1过表达TAKI则抑制SGC-7901细胞凋亡(P0.01)。此外,过表达miR-146a和敲低TAK1均能显著上调NF-κB抑制蛋白α(IκBα)和下调B细胞淋巴瘤-2(Bcl-2)表达。结论:本研究结果表明miR-146a通过靶向TAK1抑制NF-κB途径,从而诱导胃癌细胞凋亡。
[Abstract]:Aim: to observe the effect of minimal RNA-146a miR-146a on apoptosis of gastric cancer SGC-7901 cells and its possible molecular mechanism. Methods: miR-146a mimicic (up-regulated miR-146a expression) and miR-146a inhibitor (down-regulated miR-146a expression) were transfected into human gastric cancer SGC-7901 cells, respectively. The expression level of miR-146a in gastric cancer cells after transfection was detected by RT-PCR PCR and CCK-8 assay and flow cytometry were used to detect the effect of miR-146a on the growth and apoptosis of SGC-7901 gastric cancer cells. RT-q PCR and Western blot assay were used to detect the effect of miR-146a on the growth and apoptosis of SGC-7901 gastric cancer cells. The effect of up-regulation or down-regulation of miR-146a on the NF-魏 B pathway of transforming growth factor- 尾 -activated kinase (1(TAK1)/) was measured. Results: in SGC-7901 cells, Upregulation of miR-146a expression significantly promoted cell apoptosis, while down-regulation of miR-146a expression significantly inhibited cell apoptosis. Compared with NC group, the expression of mRNA and protein in SGC-7901 cells were significantly decreased after transfection of SGC-7901 cells with pmiR-146a mimics. In miR-146a inhibitors transfection group, the expression of mRNA and protein in TAK1 increased significantly, suggesting that miR-146a negatively regulated TAK1 expression. Knockdown of TAK1 promoted SGC-7901 cell apoptosis (P0.01), while TAK1 overexpression TAKI inhibited SGC-7901 cell apoptosis (P0.01). Both overexpression of miR-146a and knockout of TAK1 could significantly up-regulate the expression of NF- 魏 B inhibitory protein 伪-魏 B 伪 and down-regulate the expression of B-cell lymphoma -2pBcl-2.Conclusion: this study suggests that miR-146a inhibits NF- 魏 B pathway by targeting TAK1 and thus induces apoptosis in gastric cancer cells.
【作者单位】：温州医科大学附属第二医院肝胆外科;
【基金】：温州市科技计划项目(No.Y20130231)
【分类号】：R735.2

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