CAJ | 학술논문

为研究细胞色素氧化酶CYP3A39的生物学活性，采用RT PCR方法从巴马小型猪肝组织中克隆CY P3A 39基因，与真核表达载体pcDNA3．1连接，构建重组质粒，转染 HepG2细胞，采用G418筛选，以获得稳定表达该基因的细胞株；以CY P3A特异性代谢底物睾酮为探针药物，在体外进行孵育，高效液相色谱仪测定睾酮羟基化产物6β羟基睾酮（6β OHT ）的产量。结果显示，成功克隆了CY P3A 39基因，建立了稳定表达 CY P3A 39基因的 HepG2细胞株，且其具有睾酮代谢活性。
위연구세포색소양화매CYP3A39적생물학활성，채용RT PCR방법종파마소형저간조직중극륭CY P3A 39기인，여진핵표체재체pcDNA3．1련접，구건중조질립，전염 HepG2세포，채용G418사선，이획득은정표체해기인적세포주；이CY P3A특이성대사저물고동위탐침약물，재체외진행부육，고효액상색보의측정고동간기화산물6β간기고동（6β OHT ）적산량。결과현시，성공극륭료CY P3A 39기인，건립료은정표체 CY P3A 39기인적 HepG2세포주，차기구유고동대사활성。
The CY P3A39 gene was cloned from the liver tissues of Bama miniature pig by RT-PCR ,ligated to the expression vector pcDNA3 .1 ,and then transfected into HepG2 cells .The HepG2-CYP3A39 cell line ,stably expressing CY P3A39 was established under the selection of gentamycin (G418) .The probe drug testosterone (TST) specific to CYP3A was used to incubate with the cell lines of HepG2-pcDNA3 .1 and HepG2-CYP3A39 in optimal conditions ,and the high performance liquid chromatography (HPLC) was utilized to detect the metabolites after the end of incubation .The results showed that the cell line expressing CY P3A39 was successfully estab-lished and the expressed CY P3A 39 had significantly higher metabolic activity of testosterone than that of negative control .