Constitutive expression of HGF modulates renal epithelial cell phenotype and induces c-met and fibronectin expression.
Hepatocyte growth factor (HGF) is a potent renotropic factor that has been shown to play important roles in kidney development and recovery from acute renal injury. To examine the effects of HGF on renal tubular epithelium, we generated HGF-producing renal epithelial cells by stably transfecting mIMCD-3 and OK cells with an expression plasmid containing human HGF cDNA. Expression of HGF in the transfected cells was confirmed by detection of HGF mRNA by Northern blot analysis and detection of HGF secretion into the conditioned medium by ELISA. HGF-transfected cells exhibited fibroblast-like scattered morphology and increased cell motility. They formed branching tubules when grown in 3-D collagen gel. In addition, HGF-producing cells grew faster than their parental cells, but failed to form colonies in soft agar. These phenotypic changes were inhibited by a specific, neutralizing anti-HGF antibody. Interestingly, both c-met transcript and c-met protein were increased in HGF-transfected cells, suggesting that HGF amplifies its own action via stimulation of c-met expression. Autocrine expression of HGF and c-met in renal epithelial cells also stimulated fibronectin gene expression, which was totally blocked by incubation with a neutralizing anti-HGF but not a pan-specific anti-TGF-beta antibody, suggesting that it is independent of TGF-beta production. Our data demonstrate that HGF as a single factor stimulates renal epithelial cell proliferation, migration, differentiation, and extracellular matrix remodeling, making it uniquely suited to promote renal tubulogenesis during development, as well as to reconstitute tubular integrity following acute injury.
Department of Medicine, Rhode Island Hospital, Brown University School of Medicine, Providence 02903, USA.
Y
Y
Liu
J N
JN
Centracchio
L
L
Lin
A M
AM
Sun
L D
LD
Dworkin
D000818:Animals; D002453:Cell Cycle; D002454:Cell Differentiation; D002455:Cell Division; D002465:Cell Movement; D002478:Cells, Cultured; D004847:Epithelial Cells; D005353:Fibronectins / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D017228:Hepatocyte Growth Factor / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007679:Kidney Medulla; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D009893:Opossums; D010641:Phenotype; D019859:Proto-Oncogene Proteins c-met / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D016212:Transforming Growth Factor beta / Q000502:physiology
D005353:Fibronectins; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D017228:Hepatocyte Growth Factor; D019859:Proto-Oncogene Proteins c-met