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人剪切修复基因XPD对肝癌细胞生长及ERG基因的影响

邹颖颖1,张吉翔2   

  1. 1. 江西省南昌市民德路1号 南昌大学第二附属医院消化内科
    2. 南昌大学第二附医院消化内科
  • 收稿日期:2012-01-05 修回日期:2012-05-06 出版日期:2013-01-15 发布日期:2013-01-15
  • 通讯作者: 邹颖颖

The Role of XPD in The Hepatoma Cells Growth and Its Relationship with ERG Gene

Ying-Ying ZOU 2   

  • Received:2012-01-05 Revised:2012-05-06 Published:2013-01-15 Online:2013-01-15
  • Contact: Ying-Ying ZOU

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摘要：

目的探讨人剪切修复基因XPD转染人肝癌细胞SMMC-7721后对细胞自身生长及癌基因ERG表达的影响。方法将XPD基因通过LipofectamineTM2000转染入人肝癌细胞SMMC-7721。实验分为4组,分别为重组质粒转染细胞SMMC-7721-pEGFP-N2-XPD组(XPD组)、空载质粒转染细胞SMMC-7721-pEGFP-N2组(N2组)、脂质体转染细胞SMMC-7721组(脂质体组)、肝癌细胞SMMC-7721无转染空白对照组(空白对照组)。分别用逆转录聚合酶链反应(RT-PCR)和Westernblot法检测各组细胞中XPD、ERG的mRNA和蛋白质的表达量,用四甲基偶氮唑盐比色法(MTT)检测各组细胞的增殖活力及流式细胞仪检测各组细胞的凋亡情况。结果与其他3组比较,XPD组中的ERGmRNA及蛋白表达量显著降低,而XPDmRNA及蛋白表达量明显升高(P<0.01)。转染了XPD的肝癌细胞SMMC-7721,其细胞增殖活性(0.455±0.009)显著降低,细胞凋亡率(42.06±0.01)%明显升高(P<0.001)。结论 XPD基因可以抑制癌基因ERG的表达,明显降低肝癌细胞的增殖活力并提高肝癌细胞的凋亡率。

关键词: 肝肿瘤, 转染, XPD基因, ERG基因, p53基因, 增殖, 凋亡

Abstract: Abstract Objective：To investigate the effects of transfected XPD gene on the growth of hepatoma cell SMMC-7721 and the expression of ERG gene. Methods: The XPD gene was transfected into SMMC-7721 with Lipofectamine 2000. There were four groups in this study including SMMC-7721 - pEGFP- N2-XPD group（XPD group), SMMC-7721-pEGFP-N2 group (N2 group), Lipofectamine group (Lip group ) and blank control group. Reverse transcriptase PCR and Western blot were used to measure the expressions of XPD and ERG at mRNA and protein levels,respectively.The cell prolifera tion was assessed by MTT assay and the cell apoptosis was examined by flow cytometry . Results: Compared with N2 group, Lip group and blank control group, the expression of EGR mRNA and protein decreased significantly in XPD group (P < 0.01), in contranst the expression of XPD mRNA and protein were enhanced obviously in XPD group (P < 0.01). The proliferation of SMMC-7721 cells was markedly inhibited and the apoptosis of SMMC-7721 cells was increased obviously after transfection of XPD(P < 0.01). Conclusion: The wild-type XPD could inhibit the expression of ERG ，decrease the proliferation of SMMC-7721 cells and enhanced the apoptosis of SMMC-7721 cells in vitro.

Key words: liver neoplasms, transfection, XPD gene, ERG gene, p53 gene, proliferation, apoptosis