Expression of eNOS protein and NO in human embryonic kidney 293 T cells infected with mutant Cav-1 gene
CHEN Hai-ying
WANG Lei
WANG Lan-hua
XIA Peng
ZHANG Xiao
XUE Yu-zeng
CHEN Shuang-feng
WANG Le-xin
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.
Keywords：Caveolin-1endothelial nitric oxide synthasenitric oxidelentivirus vector
Publication Date：2014-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 5-7 )