Induction of apoptosis by hepatocyte growth factor/scatter factor and its augmentation by phorbol esters in Meth A cells

Biochem Biophys Res Commun. 1998 Apr 7;245(1):278-83. doi: 10.1006/bbrc.1998.8416.

Abstract

Hepatocyte growth factor/scatter factor (HGF/SF) is a multifunctional cytokine with mitogenic, motogenic, and morphogenic activities. In addition, HGF/SF inhibits the proliferation of some tumor cell lines, but its mechanism remains poorly understood. We determined in this study whether HGF/SF induces cell death of a Meth A mouse sarcoma cell line in vitro, whose proliferation is remarkably suppressed by HGF/SF. Inhibition of Meth A cell growth by HGF/SF was dose-dependent and maximal at a concentration of 30 ng/ml. The percentage of dead cells increased to 22% upon treatment with 30 ng/ml of HGF/SF for 96 h, whereas that in untreated cultures was less than 5%. Staining of these cells nuclei with Hoechst 33342 revealed condensation of the chromatin and nuclear fragmentation. Gel electrophoresis of DNA from HGF/SF-treated cells showed a typical ladder pattern. Cells with a fractional DNA content also increased five-fold in the HGF/SF-treated cultures as analyzed by flow cytometry after propidium iodide staining. These are features typical of apoptosis. Concurrent addition of 12-O-tetradecanoylphorbol 13-acetate (TPA) with HGF/SF augmented the apoptosis induced by the growth factor, while TPA alone caused little death. This enhancement was largely blocked by addition of the specific protein kinase C inhibitor GF 109203X. These results indicate that HGF/SF induced the apoptotic cell death of the Meth A sarcoma cell line and that protein kinase C activation augmented the growth factor-induced apoptosis.

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Benzimidazoles / metabolism
  • Cell Cycle / physiology
  • Cell Survival / drug effects
  • DNA Fragmentation / drug effects
  • DNA Replication / drug effects
  • Flow Cytometry
  • Fluorescent Dyes / metabolism
  • Hepatocyte Growth Factor / pharmacology*
  • Indoles / pharmacology
  • Maleimides / pharmacology
  • Mice
  • Protein Kinase C / antagonists & inhibitors
  • Tetradecanoylphorbol Acetate / pharmacology*
  • Tumor Cells, Cultured

Substances

  • Benzimidazoles
  • Fluorescent Dyes
  • Indoles
  • Maleimides
  • Hepatocyte Growth Factor
  • Protein Kinase C
  • bisindolylmaleimide I
  • Tetradecanoylphorbol Acetate
  • bisbenzimide ethoxide trihydrochloride