Src family kinases and nitric oxide production are required for hepatocyte growth factor-stimulated endothelial cell growth.
Hepatocyte growth factor (HGF) is a potent mitogen for vascular endothelial cells (EC); however, signal transduction pathways for HGF-stimulated EC growth remain unclear. In the present study we investigated the role of Src family kinases and nitric oxide (NO) in HGF-stimulated EC growth. Human umbilical vein endothelial cells (HUVEC) were stimulated with HGF and NO was measured by an NOx analyzing HPLC system. Activation of ERK1/2 and p38 MAPK was assessed by Western blot. NO production in HUVEC increased 1.8-fold by HGF. A Src family kinases inhibitor PP1 inhibited HGF-stimulated NO production by 71%. HUVEC growth increased 1.9-fold in cell number by HGF. PP1 and Nitro-L-arginine methylester (L-NAME) inhibited HGF-stimulated HUVEC growth by 51 and by 71%. ERK1/2 and p38 MAPK were phosphorylated by HGF and a MEK inhibitor PD98059 and a p38 MAPK inhibitor SB203580 inhibited HGF-stimulated HUVEC growth by 66% and by 58%; however, HGF-induced phosphorylation of ERK1/2 and p38 MAPK was not inhibited by L-NAME, indicating that NO is not an upstream activator of ERK1/2 and p38 MAPK. These findings demonstrated that Src family kinases regulate HGF-stimulated NO production in HUVEC and that HGF stimulates HUVEC growth through NO-dependent and NO-independent pathways.
Maejima
Yasuhiro
Y
Department of Internal Medicine, Omiya Medical Center, Jichi Medical School, Amanuma-Cho 1-847, Saitama City 330-8503, Japan.
Ueba
Hiroto
H
Kuroki
Masatoshi
M
Yasu
Takanori
T
Hashimoto
Shigemasa
S
Nabata
Aoi
A
Kobayashi
Nobuhiko
N
Ikeda
Nahoko
N
Saito
Muneyasu
M
Kawakami
Masanobu
M
D000704:Analysis of Variance; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D006801:Humans; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism*; D011336:Probability; D012680:Sensitivity and Specificity; D014471:Umbilical Veins / Q000166:cytology; D019061:src-Family Kinases / Q000032:analysis / Q000378:metabolism*
D009569:Nitric Oxide; D017228:Hepatocyte Growth Factor; D019061:src-Family Kinases