The antifibrogenic effect of hepatocyte growth factor (HGF) on renal tubular (HK-2) cells is dependent on cell growth

Ciro Esposito, Bina Parrilla, Flavia Cornacchia, Fabrizio Grosjean, Filippo Mangione, Nicoletta Serpieri, Rossella Valentino, Luigi Villa, Mariarosa Arra, Vittoria Esposito, Antonio Dal Canton

Research output: Contribution to journalArticle

Abstract

Although several reports suggest an antifibrogenic effect of hepatocyte growth factor (HGF), an increased deposition of matrix induced by HGF has also been reported. These conflicting effects could result from a diverse proliferative state of the target cells. Aim of the present study was to evaluate HGF effects on growth arrested (quiescent) and actively proliferating renal tubular epithelial (HK-2) cells. HK-2 cells were cultured in RPMI medium either on agarose gel or on plastic surface in order to inhibit or to allow cell proliferation. Cells were incubated with RPMI containing HGF (50 ng/ml) for 24 h at 37°C. Untreated HK-2 were used as control. After 24 h of incubation, cells were counted by Coulter counter. (α2)IV collagen, transforming growth factor-β (TGF-β), Tissue inhibitor of metalloproteases (TIMP1 and 2) mRNA levels were determined by RT-PCR. The production of type IV collagen, c-met, proliferating cell nuclear antigen (PCNA), and SnoN, a transcriptional Smad corepressor and thus a TGF-β inhibitor, was evaluated by ELISA or western blotting. MMP-9 and 2 gelatinolytic activity was studied by zymography. Treatment with HGF did not increase HK-2 cell number and PCNA synthesis when the cells were grown on agarose as it did for cells grown on plastic surface. HGF increased (α2)IV collagen in proliferating cells whereas it reduced (α2)IV collagen and c-met synthesis in growth arrested cells. HGF treatment increased TGF-β and TIMP-2 in proliferating cells while reduced TIMP-1 mRNA levels of quiescent cells. Furthermore, production of the co repressor SnoN was significantly decreased by HGF in proliferating cells. Quiescent and proliferating HK-2 showed a different pattern of metalloproteases activity with a prevalence of MMP2 in quiescent and MMP9 in proliferating cells. In summary, HGF showed opposite effects on growth arrested and proliferating HK-2 cells favouring matrix deposition in the latter with increasing expression of collagen, TIMP-1 and TGF-β. Our results demonstrate that the proliferative state of target cells may influence the effects of HGF on extracellular matrix turnover in HK-2 cells.

Original languageEnglish
Pages (from-to)173-180
Number of pages8
JournalGrowth Factors
Volume27
Issue number3
DOIs
Publication statusPublished - 2009

Keywords

  • Cell proliferation
  • Fibrosis
  • Growth factor
  • TGF-β

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Endocrinology
  • Cell Biology
  • Medicine(all)

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