山东医药Issue(35)：5-7,3.DOI:10.3969/j.issn.1002-266X.2014.35.002
感染突变Cav-1基因的人胚肾293T细胞eNOS蛋白、NO表达变化
Expression of eNOS protein and NO in human embryonic kidney 293 T cells infected with mutant Cav-1 gene
摘要
Abstract
Objective To investigate the expression of nitric oxide synthase ( eNOS) protein and nitric oxide ( NO) in human embryonic kidney 293T cells infected with mutant Caveolin-1 (Cav-1) gene.Methods F92-Cav-1 gene was ampli-fied by PCR and inserted into the lentiviral vector backbone pLVX-mCMV-mCherry.After the recombinant plasmid trans-formation, positive clones were identified by sequencing and restriction enzyme digestion.pLVX-F92A-Cav-1-mCMV-mCherry was successfully cloned into lentiviral vector and named as F92A-Cav-1.293T cells were randomly divided into normal control group, negative control group ( empty vector group) and LV-F92A-Cav 1 group.Normal control groups were not transfected, the empty vector groups were transfected empty plasmid, LV-F92A-Cav-1 groups were transfected with LV-F92A-Cav-1.The expression of eNOS protein and NO in 293T cells infected with LV-F92A-Cav-1 plasmid were investigated by immunofluorescence and NO fluorescent probe respectively.The cell viability was measured by CCK-8.Results Com-pared with normal control group and empty vector group, the F92A-Cav-1 could increase eNOS protein expression and NO production.The Optical Density ( OD) values in the normal control groups, empty vector groups and LV-F92A-Cav-1 groups were respectively 2.233, 2.184, 2.231, pairwise comparison in three groups, all P>0.05.Conclusion The ex-pression of eNOS protein and NO are increased in 293T cells infected with mutant Cav-1.关键词
窖蛋白-1/内皮型一氧化氮合酶/一氧化氮/慢病毒载体Key words
Caveolin-1/endothelial nitric oxide synthase/nitric oxide/lentivirus vector分类
医药卫生引用本文复制引用
陈海英,汪磊,王兰花,夏鹏,张霄,薛玉增,陈双峰,王乐信..感染突变Cav-1基因的人胚肾293T细胞eNOS蛋白、NO表达变化[J].山东医药,2014,(35):5-7,3.基金项目
国家自然科学基金资助项目（81270104）；山东省自然科学基金资助项目（ZR2013HL015）。 （）
