[Study of the inhibitory effect of NF-kappaB decoy on tissue factor gene expression and FVII activation in cultured human umbilical vein endothelial cells].
OBJECTIVE
To investigate the inhibitory effect of NF-kappaB decoy on tissue factor (TF) expression and FVII activation in cultured human umbilical vein endothelial cells (HUVEC), and to explore new methods for prevention and treatment of coronary heart disease.


METHODS
NF-kappaB decoy transfection efficiency was detected by flow cytometry, NF-kappaB decoy's mechanism was analyzed by electrophoretic mobility shift assay (EMSA), TF mRNA was detected by RT-PCR, TF antigen expression on the surface of HUVEC by flow cytometry, FVIIa level in plasma incubated with HUVEC stimulated by TNF-alpha by rsTF one stage clotting method.


RESULTS
NF-kappaB decoy could be successfully transfected into HUVEC. It could compete with the endogenous kappaB cis sequence element in the regulatory regions of TF promoter to bind transcriptional factor NF-kappaB. It could also significantly inhibit the TF mRNA, TF antigen expression on the cell surface and TF function leading to activation of FVII.


CONCLUSIONS
NF-kappaB decoy could inhibit TF gene expression and FVII activation in cultured HUVEC and might be a potential new strategy for prevention and treatment of coronary heart disease.
Wang
Lin-lin
LL
Institute of Hematology, Tongji Medical College, Huazhong Science & Technology University, Wuhan 430022, China.
Wei
Wen-ning
WN
Hu
Yu
Y
Song
Shan-jun
SJ
D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005167:Factor VII / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D009838:Oligodeoxyribonucleotides / Q000235:genetics / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013925:Thromboplastin / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014471:Umbilical Veins / Q000166:cytology
C468119:NF-kappaB decoy; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D005167:Factor VII; D013925:Thromboplastin