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Effect of silencing decoy receptor 3 on biological features of hepatoma cells
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- Author:
- No author available
- Journal Title:
- World Chinese Journal of Digestology
- Issue:
- 3
- DOI:
- 10.11569/wcjd.v25.i3.234
- Key Word:
- 肝癌;诱骗受体3;LV-shDcR3;细胞增殖;细胞凋亡;Hepatoma;Decoy receptor 3;LV-shDcR3;Cell proliferation;Cell apoptosis
Abstract： 目的 了解诱骗受体3(decoy receptor 3,DcR3)在肝癌细胞中的表达水平,探讨其对肝癌细胞增殖、凋亡的影响并探讨其分子机制.方法 采用real-time PCR、Western blot以及ELISA等方法检测肝癌细胞HepG2和Huh-7与正常肝细胞HL-7702和Chang liver中DcR3 mRNA和蛋白的表达以及分泌水平.设计和构建DcR3小干扰RNA重组慢病毒(LV-shD cR3),感染HepG2和Huh-7细胞,同时以空病毒载体作为对照,Western blot验证沉默效果.感染后通过CCK-8和平板克隆形成实验观察细胞增殖情况;通过流式细胞术观察细胞凋亡情况;通过Western blot检测凋亡相关蛋白如剪切型多聚腺苷二磷酸核糖聚合酶的表达情况.通过Western blot方法检测转染前后FasL、LIGHT和TRAIL等死亡配体的表达情况.组间比较采用单因素方差分析.结果 肝癌细胞HepG2和Huh-7中DcR3的mRNA和蛋白表达水平与正常肝细胞相比明显升高,差异具有统计学意义.细胞上清中DcR3的分泌水平也明显升高.与空白组和感染对照组相比,感染LV-shDcR3组,肝癌细胞中DcR3的表达显著降低,细胞生存率明显降低,早期凋亡细胞比例明显升高,凋亡相关蛋白表达水平明显升高.与感染前相比,肝癌细胞感染LV-shDcR3后,TRAIL和FasL的表达水平明显升高,结论 DcR3在肝癌细胞中高表达,下调肝癌细胞中DcR3的表达可以抑制细胞的增殖,促进细胞凋亡,其机制可能与TRAIL和FasL凋亡通路有关.
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