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Kinetics of hepatocyte growth factor‐induced DNA synthesis in vitro: a comparative study with epidermal growth factor and insulin

Objective To describe the kinetics of in vitro hepatocyte proliferation induced by a range of doses of hepatocyte growth factor (HGF) compared with those seen with epidermal growth factor (EGF). Hepatocyte growth factor with/without insulin, or EGF with insulin, were added to primary rat hepatocytes 20 h after plating. Methods The cumulative fraction of cells with nuclear labelling following the addition of 3H-thymidine was determined by autoradiography at intervals from 32–66 h. Results Several cell cycle parameters were determined. Hepatocyte growth factor and ECF were shown to have identical pre-replicative lag times (17 h) at all doses. Entry of hepatocytes into DNA synthesis after stimulation with HGF obeyed first order kinetics, as does entry after EGF stimulation. The maximal rate constant for the rate of entry into DNA synthesis (kG1/s) was significantly greater for HGF (0.0229/h) and HGF with insulin (0.0275/h) than EGF with insulin (0.0194/h; P < 0.005). Conclusion Hepatocyte growth factor acts as a strong proliferative stimulus at the G1 to 5 interface, compatible with the hypothesis that it is a major physiological stimulus to liver regeneration.