2006•Zhongguo jiceng yiyaoRequires access

Small interfering RNA mediated silencing of cytochrome P450 3A4 gene

Chang He, 赵立子, 邓颖, 王雪丁, 关溯, 黄民

Abstract

目的研究质粒表达型短发夹RNA(small hairpin RNA,shRNA)对CHL-3A4转基因细胞的细胞色素P450 3A4(cytochrome P450 3A4,CYP3A4)基因表达的影响.方法设计并构建3条shRNA真核表达载体(CYP3A4Ⅰ、CYP3A4Ⅱ、CYP3A4Ⅲ),脂质体转染CHL-3A4转基因细胞.转染48 h后,TR-PCR和Western Blotting分别观察3条shRNA对CYP3A4mRNA和蛋白表达的影响;噻唑蓝(MTT)观察shRNA抑制环磷酰胺对CHL-3A4转基因细胞毒性作用.结果 CYP3A4Ⅲ表达载体的shRNA分别能抑制CHL-3A4细胞的CYP3A4mRNA表达(70%)及蛋白表达(75%),抑制环磷酰胺对CHL-3A4细胞(75%)细胞毒作用.结论 RNA干扰能显著抑制CYP3A4基因的表达,RNA干扰技术为肝细胞色素P450的研究提供了新的方法。

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Key concepts: Medicine, Small interfering RNA, Gene silencing, CYP3A4, Cytochrome P450, RNA, Trans-acting siRNA, Gene