中文摘要：

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。

英文摘要：

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 pUC19 to get pU6-MGMTi, co-transfected with pEGFP-C1 into 16HBE 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. Transfected 16HBE 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.