| RNA干扰诱导DNA修复基因hMGMT表达沉寂的研究 |
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| 引用本文： | 赖延东,吕嘉春,李秀英,宾晓农,吴建军. RNA干扰诱导DNA修复基因hMGMT表达沉寂的研究[J]. 癌变．畸变．突变, 2006, 18(1): 9-11 |
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| 作者姓名： | 赖延东 吕嘉春 李秀英 宾晓农 吴建军 |
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| 作者单位： | 广州医学院化学致癌研究所,广东,广州,510182;暨南大学药学院基因组药物研究所,广东,广州,510632 |
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| 基金项目： | 国家自然科学基金项目（No.30371196,30200235）;广东省重点科技攻关资助项目（No.2002830104）;广东省医学科研资助项目（B20001075）. |
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| 摘 要： | 背景与目的：构建能特异诱导人DNA修复基因hMGMT表达沉寂的RNA干扰载体。材料与方法：通过两步法聚合酶链反应（Polymerasing chain reaction，PCR）扩增hMGMT特异RNA干扰表达盒，连入质粒载体构建RNA干扰质粒pU6-MGMTi，与pGEFP-C1共转染人支气管上皮16HBE（Human bronchial epithelial）细胞，G418筛选后，采用RT-PCR和Westem Blot分别检测mRNA和蛋白水平的表达抑制情况。结果：成功构建hMGMT特异RNA干扰质粒pU6-MGMTi，转染16HBE细胞的mRNA和蛋白表达受到显著的抑制。结论：应用RNA干扰表达盒以及共转染技术，构建MGMT基因表达沉寂的细胞株，为进一步阐明MGMT基因的功能创造条件。
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| 关 键 词： | hMGMT基因 RNA干扰 16HBE |
| 文章编号： | 1004-616X（2006）01-0009-03 |
| 收稿时间： | 2005-03-10 |
| 修稿时间： | 2005-03-102005-05-20 |
Study of RNA Interference Induced Expression Silencing of DNA Repair Gene hMGMT |
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| LAI Yan-dong,LU Jia-chun,LI Xiu-ying,BIN Xiao-nong,WU Jian-jun. Study of RNA Interference Induced Expression Silencing of DNA Repair Gene hMGMT[J]. Carcinogenesis,Teratogenesis and Mutagenesis, 2006, 18(1): 9-11 |
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| Authors: | LAI Yan-dong LU Jia-chun LI Xiu-ying BIN Xiao-nong WU Jian-jun |
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| Affiliation: | Institute for Chemical Carcinogenesis, Guangzhou Medical College, Guangzhou 510182, Guangdong , China; 2. Institute for Genome Medicine Research of School of Pharmaceutical, Jinan University 510632, Guangzhou, China |
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| Abstract: | BACKGROUND&AIM: To construct the RNA interference vector which can specially induce the expression silencing of human DNA repair gene hMGMT. MATERIAL AND METHODS: The hMGMT specific siRNA expression cassette was made by two steps PCR,and then linked with pUC19to get pU6-MGMTi,co-transfected with pEGFP-C1into16HBE and screened by G418.The expression level of mRNA and protein was detected by RT-PCR and Western Blot,respectively. RESULTS: hMGMT-specific RNA interference vector pU6-MGMTi was constructed successfully.Transfected16HBE cells MGMT mRNA and protein expression level were dramatically suppressed. CONCLUSION: Silencing of MGMT expression cell line was built by RNA interference expression cassette and co-transfection technology,which offered condition for studying the gene function of MGMT. |
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| Keywords: | hMGMTgene RNA interference 16HBE |
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