期刊论文

中文

癌变·畸变·突变

1004-616X

2018

30

2

126-131

10.3969/j.issn.1004-616x.2018.02.009

深圳市科技研发基础研究项目

本校为其他机构

目的:构建类固醇合成急性调节蛋白(StAR)基因高表达载体,建立StAR基因高表达细胞,研究StAR基因高表达对邻苯二甲酸二-(2-乙基己)酯(DEHP)毒性作用的影响。方法:根据GenBank提供的StAR基因cDNA序列设计引物,PCR扩增StAR基因并将其克隆到慢病毒载体中,构建StAR基因高表达MCF-7细胞,通过基因测序、荧光定量PCR和Western blot鉴定细胞。用不同剂量DEHP染毒MCF-7细胞和StAR基因高表达MCF-7细胞24 h,检测细胞凋亡相关基因Bax、Caspase-3、Caspase-8在mRNA和蛋白表达水平的变化。结果:基因测序证明构建的StAR基因高表达载体序列正确,荧光定量PCR检测StAR基因转染细胞比正常MCF-7细胞StAR mRNA表达水平升...

OBJECTIVE:To construct cells with over-expression of the StAR gene and to study its apoptotic response to di-(2-ethylhexyl) phthalate (DEHP). METHODS:Primers were designed according to cDNA sequence of the StAR gene from GenBank. The gene was amplified using PCR and ligated into the lentiviral vector pLVX-CMV- ZSgreen-PGK-Puro. 293FT cells were transfected with the recombinant vector and then MCF-7 cells were transfected. Cells with over-expression of the StAR gene were identified by gene sequencing,real-time quantitative PCR and western blot. Then the StAR gene over-expression cells and MCF-7...