| HBx基因缺失突变体HBx-d382对L02细胞增殖及非锚定依赖生长能力的影响 |
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| 引用本文： | 胡志亮,谭德明,侯周华,谢萍,刘国珍,欧阳奕,刘菲,刘洪波.HBx基因缺失突变体HBx-d382对L02细胞增殖及非锚定依赖生长能力的影响[J].世界华人消化杂志,2010,18(11). |
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| 作者姓名： | 胡志亮 谭德明 侯周华 谢萍 刘国珍 欧阳奕 刘菲 刘洪波 |
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| 作者单位： | 中南大学附属湘雅医院传染科,湖南省长沙市,410087 |
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| 摘 要： | 目的:建立稳定表达HBx基因缺失突变体(HBx-d382)的L02肝细胞株,并探讨其对L02细胞增殖的影响.方法:含HBx-d382重组质粒(pCDNA3.0/HBx-d382)经过PCR扩增、双酶切及测序鉴定后,通过脂质体转染和G418筛选获得稳定表达HBx-d382的L02肝细胞株.PCR鉴定基因组中HBx-d382基因整合.RT-PCR和Western blot鉴定其表达,进一步通过MTT法,软琼脂克隆形成实验检测HBx-d382对L02细胞增殖及非锚定依赖生长能力的影响.用流式细胞仪检测其对细胞周期的影响.结果:经PCR扩增、双酶切及测序鉴定HBx-d382重组质粒构建正确.稳定转染该质粒的L02细胞基因组存在HBx-d382整合.RT-PCI汲Western blot表明在RNA水平和蛋白水平存在HBx-d382表达,稳定表达HBx-d382的L02细胞其增殖能力和非锚定生长能力增强,S+G2期细胞比例升高.结论:成功构建了HBx缺失突变体的真核表达模型,证实HBx缺失突变体能影响细胞增殖,这种效应可能与其影响细胞周期调控相关.
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| 关 键 词： | 突变 增殖 细胞周期 |
HBx gene deletion mutant (HBx-d382) enhances the proliferation and anchorageindependent growth of L02 cells |
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| Zhi-Liang Hu,De-Ming Tan,Zhou-Hua Hou,Ping Xie,Guo-Zhen Liu,Yi Ouyang,Fei Liu,Hong-Bo Liu.HBx gene deletion mutant (HBx-d382) enhances the proliferation and anchorageindependent growth of L02 cells[J].World Chinese Journal of Digestology,2010,18(11). |
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| Authors: | Zhi-Liang Hu De-Ming Tan Zhou-Hua Hou Ping Xie Guo-Zhen Liu Yi Ouyang Fei Liu Hong-Bo Liu |
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| Institution: | Zhi-Liang Hu,De-Ming Tan,Zhou-Hua Hou,Ping Xie,Guo-Zhen Liu,Yi Ouyang,Fei Liu,Hong-Bo Liu,Department of Infectious Diseases,Xiangya Hospital,Central South University,Changsha 410087,Hunan Province,China |
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| Abstract: | AIM: To establish a L02 cell line stably expressing HBx gene deletion mutant (HBx-d382) and to examine their proliferation changes. METHODS: A recombinant plasmid encoding the HBx deletion mutant (pcDNA3.0/HBx-d382) was verified by PCR amplification, double restriction digestion and DNA sequencing, and then introduced into L02 cells by liposome-mediated transfection. Positive clones were selected in the present of G418. The genome integration of the HBx gene deletion mutant was confirmed by PCRamplification... |
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| Keywords: | HBx |
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