Hepatocyte growth factor stimulates the differentiation of human tracheal epithelia in vitro

Abstract

Hepatocyte growth factor (HGF) can influence epithelial cell growth and differentiation. We examined the actions of recombinant human HGF (rhHGF) on the differentiation of human primary tracheal epithelial (HTE) cells cultured in vitro for up to 96 h. Basolateral, but not apical, treatment of confluent HTE cell sheets for 48 h with rhHGF led to increases in cell height, cell volume, cilia, and total protein content. Basolateral rhHGF treatment produced a decrease in HGF receptor (c-met) expression but had no effect on c-met mRNA levels. HTE cell sheets treated with rhHGF for 48 h showed a significant increase in mediator-induced Cl- secretion and a decrease in amiloride-sensitive sodium absorption. No effect on transepithelial resistance was observed with rhHGF treatment. The enhancement of short-circuit responses by basolateral rhHGF was dose dependent. Our results demonstrate that rhHGF has hypertrophic actions on, and can influence the differentiation of, human airway epithelia in vitro, presumably through the activation of c-met at the basolateral surface of these cells.

Vascular Endothelial Growth Factor Governs Endothelial Nitric-oxide Synthase Expression via a KDR/Flk-1 Receptor and a Protein Kinase C Signaling Pathway

“…VEGF receptor-selective mutants Flt-sel (R82E/K84E/H86E, deficient in the KDR binding) and KDR-sel (D63A/E64A/E67A, deficient in Flt-1 binding) were prepared using the Muta-Gene Phagemid in vitro mutagenesis kit as described previously (32). The heterodimeric form of recombinant human hepatocyte growth factor (HGF) was produced in and isolated from Chinese hamster ovary cells as described previously (33). Recombinant human basic fibroblast growth factor (FGF), recombinant human placental growth factor (PlGF), recombinant human transforming growth factor (TGF)-␤1, TGF-␤2, and recombinant human epidermal growth factor (EGF) were purchased from R & D Systems, Inc. (Minneapolis, MN).…”

Homologous Up-regulation of KDR/Flk-1 Receptor Expression by Vascular Endothelial Growth Factor in Vitro

“…Cell Lysis and Western Blot Detection of KDR-The methods for cell lysis and Western blot have been described in detail elsewhere (43). A rabbit anti-KDR polyclonal antibody (Santa Cruz Biotech, Santa Cruz, CA) was used to probe for KDR protein.…”

Induction of Angiogenesis by Expression of Soluble Type II Transforming Growth Factor-β Receptor in Mouse Hepatoma

“…Multiple exposures of films were obtained to determine the optimal exposure time. The method for cell and tissue lysis and Western blot has been described previously (26). The mouse VEGF (Santa Cruz Biotechnology, Inc.), basic fibroblast growth factor (bFGF, Santa Cruz Biotechnology, Inc.), eNOS (Transduction Laboratories Corp., Lexington, KY), flk-1 (Santa Cruz Biotechnology, Inc.), and actin (Santa Cruz Biotechnology, Inc.) antibodies were used as primary antibodies.…”