Heparin-binding epidermal growth factor-like growth factor expression in cultured fetal human vascular smooth muscle cells. Induction of mRNA levels and secretion of active mitogen.
Heparin-binding EGF-like growth factor (HB-EGF) is a recently identified potent mitogen for smooth muscle cells (SMC). To explore whether SMC can also synthesize HB-EGF, cultured fetal human vascular SMC (FHVSMC) were analyzed for the production of HB-EGF mRNA and active growth factor. It was found that in FHVSMC, HB-EGF has the characteristics of an early response gene in that (i) the addition of fresh 10% fetal calf serum to serum-starved FHVSMC led to a rapid and transient rise in HB-EGF mRNA levels with a maximal induction of 12-14-fold occurring within 2-4 h, (ii) the phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA) also elevated HB-EGF mRNA levels rapidly and transiently with a maximal induction of 7-8-fold occurring at 2-4 h, and (iii) cyclohexamide at 40 micrograms/ml markedly increased basal, serum-, and TPA-induced HB-EGF mRNA levels. In addition, HB-EGF mRNA levels were increased 7-11-fold by addition of either HB-EGF itself, platelet-derived growth factor, or basic fibroblast growth factor, all potent SMC mitogens. Besides synthesizing HB-EGF mRNA, FHVSMC were found to release into conditioned medium a bioactive HB-EGF-like protein that cross-reacted with anti-HB-EGF antibody.
Dluz
S M
SM
Department of Surgical Research, Children's Hospital, Boston, Massachusetts 02115.
Higashiyama
S
S
Damm
D
D
Abraham
J A
JA
Klagsbrun
M
M
D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D016895:Culture Media, Serum-Free; D003513:Cycloheximide / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D005333:Fetus; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D015870:Gene Expression; D006133:Growth Substances / Q000494:pharmacology*; D006493:Heparin / Q000378:metabolism; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007700:Kinetics; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000302:isolation & purification; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D003470:Culture Media; D016895:Culture Media, Serum-Free; D006133:Growth Substances; C587883:HBEGF protein, human; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D010982:Platelet-Derived Growth Factor; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor; D006493:Heparin; D003513:Cycloheximide; D013755:Tetradecanoylphorbol Acetate