人5α-还原酶Ⅱ在CHO细胞中的表达及活性鉴定  

Expression and Activity Assay of Human Steroid 5 Alpha-Reductase Type Ⅱ in CHO Cells

作　　者：张健 高小平 

机构地区：[1]成都地奥制药集团药物研究所,四川成都610041

出　　处：《药物生物技术》2004年第5期281-285,共5页Pharmaceutical Biotechnology

摘　　要：类固醇5α还原酶(SRD5A)催化睾酮(T)转化为二氢睾酮(DHT),DHT的升高可导致良性前列腺增生等疾病的发生,因此抑制SRD5A的活性将成为治疗该类疾病的重要途径。文章将5α还原酶Ⅱ(SRD5A2)基因克隆到pcDNA3载体,并转染至CHO细胞,经G418筛选后获得表达SRD5A2基因的CHO细胞系。应用新建立的表达SRD5A2的细胞系,检测了选择性和非选择性抑制剂对SRD5A2的抑制效应,结果表明外源性SRD5A2显示很高的酶活性,而且这种活性能被其特异抑制剂非那雄胺所抑制。这一结果不仅确证了人类固醇5α还原酶Ⅱ型在CHO细胞的表达和产物活性,同时也为该类抑制剂的体外筛选提供了一种简便可靠的方法。Steroid 5alpha reductase (SRD5A) catalyzes the conversion of testosterone (T) to dihydrotestosterone (DHT). DHT is primarily responsible for the pathogenesis of benign prostatic hyperplasia (BPH), thus the inhibition of SRD5A activity was regarded as an effective approach for the therapy of BPH. The human steroid 5 alpha reductase type Ⅱ (hSRD5A2) was constructed by using the pcDNA3 vector and transfected into the CHO cells. By selection with G418 sulfate, a CHO cell lines expressing hSRD5A2 was obtained. The newly established cell lines was used for testing the inhibitory effects of selective and non selective inhibitor on human 5alpha reductase type Ⅱ. The results indicated that the foreign SRD5A2 have a high enzymatic activity and its activity was inhibited effectively by selective inhibitor Finasteride. Our results not only demonstrate the expression and activity of human steroid 5 alpha reductase type Ⅱ in CHO cells, but also provide a simple and effective method for screening inhibitors of steroid 5alpha reductase Ⅱ.

关 键 词：5α-还原酶Ⅱ CHO表达 前列腺增生 酶活性分析 非那雄胺 

分 类 号：Q55[生物学—生物化学]