title: Src family kinases and nitric oxide production are required for hepatocyte growth factor-stimulated endothelial cell growth.
abstract: 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.
authors.affiliation: Department of Internal Medicine, Omiya Medical Center, Jichi Medical School, Amanuma-Cho 1-847, Saitama City 330-8503, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Maejima
authors.forename: Hiroto
authors.initials: H
authors.lastname: Ueba
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Kuroki
authors.forename: Takanori
authors.initials: T
authors.lastname: Yasu
authors.forename: Shigemasa
authors.initials: S
authors.lastname: Hashimoto
authors.forename: Aoi
authors.initials: A
authors.lastname: Nabata
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Kobayashi
authors.forename: Nahoko
authors.initials: N
authors.lastname: Ikeda
authors.forename: Muneyasu
authors.initials: M
authors.lastname: Saito
authors.forename: Masanobu
authors.initials: M
authors.lastname: Kawakami
mesh_terms: 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*
chemical_list: D009569:Nitric Oxide; D017228:Hepatocyte Growth Factor; D019061:src-Family Kinases