Effects of hepatocyte growth factor on the growth and metabolism of human hepatocytes in primary culture.
The effect of recombinant human hepatocyte growth factor (h-rHGF), a potent mitogen for hepatocytes, was investigated in primary cultures of human hepatocytes. Here, we describe a series of experiments to investigate the kinetics of its mitogenic action, as well as its metabolic effects on cultured human hepatocytes. The h-rHGF is a potent signal for initiating DNA synthesis in human hepatocytes, with maximal stimulatory effects at 10 ng/mL (0.1 pmol/L). The kinetics of DNA synthesis showed a lag of about 48 to 72 hours, followed by a maximum at 96 hours. At least 48 hours of continuous exposure to h-rHGF are required to initiate DNA synthesis in quiescent human hepatocytes. Cell cycle analysis by flow cytometry showed that most of quiescent 2c cells have left G0/G1 and entered the cell cycle (S and G2/M phases) by 96 hours of continuous exposure to h-rHGF. When compared with other growth factors, h-rHGF was a much more potent mitogen. The effects of 10 ng/mL (0.1 pmol/L) h-rHGF on DNA synthesis were only achieved by 1.5 pmol/L epidermal growth factor (EGF), 0.1 mumol/L insulin, or 1 mumol/L glucagon. It is noteworthy that the effect of h-rHGF was potentiated by glucagon but not by insulin or EGF. The stimulatory effect of HGF on DNA synthesis was gradually inhibited by h-rHGF transforming growth factor beta (TGF-beta) in the range 1 to 10 ng/ml. The HGF also influenced the expression of other hepatic genes.(ABSTRACT TRUNCATED AT 250 WORDS)
Gómez-Lechón
M J
MJ
Unidad de Hepatología Experimental, Centro de Investigación, Hospital, Universitario La Fe, Valencia, Spain.
Castelli
J
J
Guillén
I
I
O'Connor
E
E
Nakamura
T
T
Fabra
R
R
Trullenque
R
R
D000210:Acute-Phase Reaction / Q000097:blood; D000328:Adult; D000368:Aged; D001798:Blood Proteins / Q000032:analysis; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D005260:Female; D006003:Glycogen / Q000378:metabolism; D006133:Growth Substances / Q000494:pharmacology; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D006728:Hormones / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D011994:Recombinant Proteins
D001798:Blood Proteins; D006133:Growth Substances; D006728:Hormones; D011994:Recombinant Proteins; D017228:Hepatocyte Growth Factor; D006003:Glycogen; D004247:DNA