eNOS基因在JAR细胞中的表达

Expression of transfected eNOS gene in human JAR cell

本研究应用脂质体介导转染技术将人内皮型一氧化氮合酶 (eNOS)基因转入绒癌细胞系JAR细胞 ,获得转染阳性细胞。用RT PCR和Westernblot技术从基因及蛋白水平对表达产物进行鉴定 ,结果显示 :有较高水平的mRNA转录和特异目的蛋白表达 ;通过免疫细胞化学方法证实 ,转染eNOS基因的阳性JAR细胞与对照组相比 ,有外源eNOS蛋白高表达 ;但是一氧化氮合酶 (NOS)活性及其催化产物NO并没有直接升高 ;使用A2 3187处理则能增加NOS的活性 ,说明在转染的JAR细胞中 。

Activity of endothelial nitric oxide synthase(eNOS)is regulated in a complex fashion by eNOS “cofactor-accumulating” enzymes as well as more than 20 interacting proteins.The present study was designed to determine the enzyme activity of eNOS in the transfected JAR cell.The results showed that mRNA of eNOS in the transfected JAR cell significantly increased compared to untreated cells.Immunocytochemistry revealed a strong green perinuclear immunofluorescence in eNOS transfected JAR cell.NOS activity and NO release did not change in transfectants.In this study,three independent methods demonstrated transfection resulted in over-expression of eNOS in JAR cell,but the enzyme was not active until treated with A23187.