microRNA-124对胃癌细胞增殖与凋亡的影响及其与PI3K/Akt信号通路的关系
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韦维, Email: ww62589@163.com

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广西教育厅高校科研基金资助项目(YB2014293)。


Effects of microRNA-124 on proliferation and apoptosis of gastric cancer cells and its relation with PI3K/Akt signaling pathway
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    摘要:

    背景与目的:研究显示,microRNA-124(miR-124)在多种肿瘤中发挥抑癌基因的功能,并且在胃癌细胞及组织中表达下调,在胃癌的发生发展中发挥重要调控作用。本研究探讨miR-124对人胃癌细胞增殖、凋亡的影响及其与PI3K/Akt信号通路的关系。
    方法:用qRT-PCR检测人胃癌细胞系MGC803、AGS、MNK-45、SGC-7901和人正常胃黏膜上皮细胞GES-1中miR-124的表达。用miR-124模拟物(miR-124模拟物组)及miR-124阴性对照序列(阴性对照组)分别转染至MGC803细胞后,用CCK-8法及细胞克隆实验检测MGC803细胞增殖情况,流式细胞术检测细胞凋亡,Western blot检测PI3K、Akt、p-PI3K和p-Akt蛋白水平,qRT-PCR检测PI3K和Akt mRNA 表达。
    结果:与人正常胃黏膜上皮细胞GES-1比较,miR-124在各胃癌细胞中的表达均明显降低(均P<0.05)。与阴性对照组比较,miR-124模拟物组转染后的OD450值与细胞克隆形成数均明显降低(P<0.05);细胞凋亡率明显增加(P<0.05);PI3K/Akt信号通路中关键分子PI3K、Akt、p-PI3K和p-Akt蛋白水平明显降低(均P<0.05),PI3K和Akt基因表达水平也明显降低(均P<0.01)。
    结论:miR-124表达的下调可促进胃癌细胞的增殖、抑制其凋亡,该作用机制可能与PI3K/Akt信号通路活化有关。

    Abstract:

    Background and Aims: Previous studies have demonstrated that microRNA-124 (miR-124) plays an important role in a variety of cancers, and is down-regulated in gastric cancer cells and tissues. This study was conducted to investigate the influences of miR-124 on proliferation and apoptosis of gastric cancer cells and its association with PI3K/Akt signaling pathway.  
    Methods: The expressions of miR-124 in human gastric cancer cells MGC803, AGS, MNK-45 and SGC-7901 as well as in normal gastric mucosa epithelial GES-1 cells were determined by qRT-PCR. In MGC803 cells after transfected with miR-124 mimics (miR-124 mimics group) and negative control sequences (negative control group), the cell proliferation was detected by CCK-8 assay and cell clone assay, the apoptosis was measured by flow cytometry, the protein expressions of PI3K, Akt, p-PI3K and p-Akt were examined by Western blot, and mRNA expressions of PI3K and Akt were analyzed by qRT-PCR. 
    Results: The expressions of miR-124 in all studied gastric cancer cell lines were all significantly lower than that in normal gastric mucosa epithelial GES-1 cells (all P<0.05). In miR-124 mimics group compared with negative control group after transfection, the OD450 values of and numbers of cell clone formation were all significantly reduced (all P<0.05); the apoptosis rates were all significantly increased (all P<0.05); the expression levels of PI3K, Akt, p-PI3K and p-Akt proteins from PI3K/Akt signaling pathway were significantly down-regulated (all P<0.05) and the gene expression levels of PI3K and Akt were also significantly down-regulated (all P<0.05). 
    Conclusion: Down-regulation of miR-124 can promote proliferation and inhibit apoptosis of gastric cancer cells, and the mechanism may probably associated with the activation of PI3K/Akt signaling pathway.

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韦维, 黄海舸, 陆佳明, 周莅. microRNA-124对胃癌细胞增殖与凋亡的影响及其与PI3K/Akt信号通路的关系[J].中国普通外科杂志,2020,29(6):723-730.
DOI:10.7659/j. issn.1005-6947.2020.06.013

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  • 收稿日期:2019-09-19
  • 最后修改日期:2020-04-21
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  • 在线发布日期: 2020-06-25