利用GADD153启动子建立替代动物试验检测致癌物的方法研究  

Alternative methods to animal experiment for carcinogen detection by GADD153 promoter

作　　者：韩喜彬[1] 王立辛[2] 巴彩凤[1] 

机构地区：[1]辽宁医学院基础医学院,辽宁锦州121001 [2]辽宁医学院畜牧兽医学院,辽宁锦州121001

出　　处：《畜牧与兽医》2014年第6期33-36,共4页Animal Husbandry & Veterinary Medicine

基　　金：辽宁省科技厅2011年计划项目(2011408004)

摘　　要：本试验旨在构建可在一定程度上代替动物试验,用于快速筛查化合物致癌性的重组SMMC-7721细胞株。方法:构建含有生长抑制与DNA损伤基因153(GADD153)启动子和绿色荧光蛋白报告基因的重组质粒,稳定转染SMMC-7721细胞;以荧光检测CdCl2对转染细胞的DNA损伤效应;同时以RT-PCR检测内源性GADD153 mRNA的表达,验证荧光强度、GADD153 mRNA表达量依CdCl2浓度变化的剂量依赖关系。结果表明:随着CdCl2浓度的增加,GADD153-EGFP细胞株的荧光强度、GADD153 mRNA表达量呈剂量依赖性增加。重组GADD153-EGFP细胞株可检测致癌物的DNA损伤效应,说明此方法代替动物实验是可能的。The aim of this study is to construct a recombinant GADD153-EGFP cell lines that could replaced the animal experiment to some extent for detection of carcinogen. A recombinant plasmid containing DNA damage and repair inducible gene 153 （GADD153） promoter and green fluorescence protein reporter gene was constructed, and then transfected into SMMC -7721 cells. DNA damage induced by CdC12 was detected by bioluminescence, and endogenic GADD153 mRNA level was detected by RT-PCR to verify the dose-response relationship of fluorescence intensity and GADD153 mRNA expression. With the increase of concentration of CdCI2, fluorescence intensity and GADD153 mRNA expression quantity increased with a dose-dependent manner in GADD153-EGFP cell lines. Recombinant GADD153-EG- FP cell could detect DNA damage effects, indicating that it was possible that this method replaced animal test.

关 键 词：替代方法 生长抑制与DNA损伤基因153 致癌物 

分 类 号：S852.2[农业科学—基础兽医学]