线粒体融合素基因-2对肝癌细胞凋亡的影响及其机制
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郑启昌

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Effect of mitofusin-2 gene on apoptosis of hepatic carcinoma cell strain and its possible mechanism
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    摘要:

    目的:探讨外源性线粒体融合素基因-2(mfn2)对肝癌细胞株HepG2凋亡的影响及其机制。
    方法:利用脂质体2000将质粒mfn2转染HepG2细胞。RT-PCR检测转染后细胞mfn2 mRNA的表达水平;Weastern Blot法检测mfn2蛋白的表达;MTT检测mfn2对HepG2细胞增殖的影响;Annnexin-V/PI双标记法检测转染前后细胞凋亡的变化;RT-PCR检测凋亡相关基因的表达;电镜观察超微结构的变化。
    结果: 转染mfn2基因的HepG2细胞可以稳定表达Mfn2蛋白;MTT实验提示转染mfn2基因后,HepG2细胞增殖受到抑制;转染组与转染空质粒组和空白对照组相比前者可促进细胞凋亡(P<0.05);电镜示mfn2转染后HepG2细胞线粒体嵴断裂、消失、基质疏松;RT-PCR结果示Bcl-2和survivin基因表达下调。
    结论:外源性mfn2基因可抑制HepG2细胞的增殖,诱导其凋亡。凋亡相关基因的表达下调可能是其诱导凋亡的机制。

    Abstract:

    Abstract:Objective:To investigate the effect of mitofusin-2 gene (mfn2) on apoptosis of hepatocellular carcinoma cell strain HepG2 and its possible mechanism.
    Methods :Plasmid pEGFPmfn2 was transfected into hepatocellular carcinoma cell strain HepG2 by Lipofectamine 2000 in vitro. The mRNA of Mfn2 expression was detected by RT-PCR and protein expression by Western-blot. The cell proliferation of HepG2 was measured by MTT assay and cell apoptosis was observed in Annexin-V/PI by flow cytometry. The ultramicrostructure of cell was observed by electron microscope. The expression of apoptosis related genes was observed by RT-PCR.
    Results:Mitofusin-2 gene was expressed in transfected HepG2/mfn2 stably and it could obviously inhibit cell proliferation by MTT assay. Exogenous mfn2 gene significantly induced cell apoptosis and induced the break down, disappearance and groundsubstance rarefaction of chondrosome cristae. Expression of Bcl-2 and survivin genes were reduced in HepG2/mfn2.
    Conclusions:Exogenous mfn2 could inhibit proliferation of HepG2 and induce its apoptosis. The downregulation of apoptosis related genes may be the mechanism for inducing apoptosis.

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王尧,郑启昌,胡文君,胡青钢,夏耘,张景辉.线粒体融合素基因-2对肝癌细胞凋亡的影响及其机制[J].中国普通外科杂志,2008,17(7):11-677.
DOI:10.7659/j. issn.1005-6947.2008.07.011

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  • 在线发布日期: 2008-07-25