期刊论文

中文

现代预防医学

1003-8507

2018

45

09

1657-1662

深圳市科技研发基础研究项目（JCYJ20170306160553495）；

本校为其他机构

目的 构建3β-HSD高表达细胞株,研究3β-HSD高表达对塑化剂DEHP致凋亡的影响.方法 根据GenBank上3β-HSD基因cDNA序列设计引物,PCR扩增该基因并将其克隆到慢病毒载体pLVX-CMV-ZSgreen-PGK-Puro,然后转染至MCF-7细胞中,对细胞株进行鉴定.用不同剂量DEHP对3β-HSD高表达细胞和MCF-7细胞染毒24h,在基因和蛋白表达水平上检测Bax、Caspase-3、Caspase-8的变化.结果 基因测序验证本文构建的3β-HSD基因高表达载体基因序列与GenBank中的序列一致,荧光定量PCR结果显示3β-HSD基因表达水平升高105倍,western blot结果显示3β-HSD蛋白表达水平升高80％.DEHP染毒MCF-7细胞后,Bax、Caspase-3、Caspase-8基因...

Objective To construct a lentiviral vector with 3β-HSD gene overexpression and transfect it into human MCF-7 cells, and to construct a cell line with 3β-HSD gene overexpression, finally to study the effects of 3β-HSD on apoptosis induced by di-(2-ethylhexyl) phthalate (DEHP). Methods Primers were designed according to cDNA sequence of 3β-HSD gene from GenBank. The gene was amplified using PCR and ligated into the lentiviral vector pLVX-C-ZSgreen-PGK-Puro.293FT cells were transfected with the recombinant vector, and MCF-7 cells were then tra...