| N-Ras基因靶向的shRNA抑制二羟环氧苯并芘诱导的恶变细胞增殖 |
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| 引用本文： | 周兰兰,蒋义国,谭爱军,沈月兰,刘林华,杨巧媛. N-Ras基因靶向的shRNA抑制二羟环氧苯并芘诱导的恶变细胞增殖[J]. 环境与健康杂志, 2009, 26(2): 101-103 |
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| 作者姓名： | 周兰兰 蒋义国 谭爱军 沈月兰 刘林华 杨巧媛 |
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| 作者单位： | 广州医学院化学致癌研究所与预防医学系,广东,广州,510182;珠海市疾病预防控制中心,广东,珠海,519000;广州医学院化学致癌研究所与预防医学系,广东,广州,510182;珠海市疾病预防控制中心,广东,珠海,519000 |
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| 基金项目： | 国家自然科学基金资助项目(30571546,30771780);;教育部留学回国人员科研启动基金资助项目(2007-24);;广东省高校自然科学重点项目(06Z021);;广东省自然科学基金资助项目(07117550) |
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| 摘 要： | 目的用小发卡RNA(small hairpin RNA,shRNA)干扰技术沉默N-Ras基因表达,探讨N-Ras基因表达对二羟环氧苯并芘(BPDE)转化人支气管上皮细胞(16HBE-T)恶性性状的影响。方法将已建立的针对N-Ras基因的shRNA干扰表达质粒载体pGPU6/GFP/Neo-NRas-566转染16HBE-T细胞,经G418筛选,建立稳定沉默N-Ras基因转化细胞系。采用Western blot筛选最佳N-Ras基因沉默细胞系,用流式检测N-Ras沉默细胞周期变化,用软琼脂实验和裸鼠成瘤实验进行细胞表型分析。结果成功培养出稳定沉默N-Ras基因的16HBE-T细胞系,Western blot显示N-Ras蛋白抑制率最高达86%。流式分析发现稳定沉默N-Ras基因表达诱导转化细胞G0/G1期捕获,软琼脂实验和裸鼠成瘤实验发现稳定沉默N-Ras基因降低了体外恶性转化细胞的集落形成率、抑制了体内肿瘤细胞生长。结论N-Ras基因沉默可以降低转化细胞的细胞周期进程和细胞恶性性状表型,提示N-Ras基因在二羟环氧苯并芘诱导细胞恶变过程中具有重要作用。
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| 关 键 词： | 二羟环氧苯并芘 RNA干扰 小发夹RNA N-Ras基因 |
shRNA Targeting N-Ras Inhibits Proliferation of Transformed Cells Induced by Anti-benzo (a)pyrene-trans-7,8-dihydrodiol-9,10-epoxide(BPDE) |
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| ZHOU Lan-lan,JIANG Yi-guo,TAN Ai-jun,et al.. shRNA Targeting N-Ras Inhibits Proliferation of Transformed Cells Induced by Anti-benzo (a)pyrene-trans-7,8-dihydrodiol-9,10-epoxide(BPDE)[J]. Journal of Environment and Health, 2009, 26(2): 101-103 |
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| Authors: | ZHOU Lan-lan JIANG Yi-guo TAN Ai-jun et al. |
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| Affiliation: | ZHOU Lan-lan,JIANG Yi-guo,TAN Ai-jun,et al.Institute for Chemical Carcinogenesis , Department of Preventive Medicine,Guangzhou Medical College,Guangzhou,Guangdong 510182,China |
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| Abstract: | Objective To explore the effects of N-Ras gene over expression on malignantly transformed human bronchial epithelial cell induced by anti-benzo(a)pyrene-trans-7,8-dihydrodiol -9,10-epoxide(BPDE)(16HBE-T) using small hairpin RNA(shRNA).Methods The recombinant plasmid pGPU6/GFP/Neo-NRas-566 including N-Ras shRNA was transfected to 16HBE-T cells.Stable N-Ras-silencing clonal 16HBE-T cell lines were selected by G418 and their N-Ras expressions were detected with Western blot to choose the clonal cell line with ... |
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