Bronchial epithelial cell growth regulation in fibroblast cocultures: the role of hepatocyte growth factor.
Proliferation of bronchial epithelial cells is an important biological process in physiological conditions and various lung diseases. The objective of this study was to determine how bronchial fibroblasts influence bronchial epithelial cell proliferation. The proliferative activity in cocultures was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and direct cells counts. Concentration of cytokines was measured in cell culture supernatants by means of ELISA. In primary cell cocultures, fibroblasts or fibroblast-conditioned medium enhanced 1.85-fold the proliferation of primary bronchial epithelial cells (P < 0.02) compared with bronchial epithelial cells cultured alone. The proliferative activity in cocultures and in fibroblast-conditioned medium was reduced by neutralizing antibody to hepatocyte growth factor (HGF) and HGF receptor c-met. Neutralizing antibodies to FGF-7 and IGF-1 had no effect. Treatment of fibroblast-epithelial cocultures with anti-IL-6 and anti-TNF-alpha neutralizing antibodies and with indomethacin decreased production of HGF. These results indicate that cytokines and PGE(2) may indirectly mediate epithelial cell proliferation via the regulation of HGF in bronchial stromal cells and that HGF plays a crucial role in proinflammatory cytokine-induced proliferation in the experimental system studied.
Skibinski
Grzegorz
G
Respiratory Medicine Research Cluster, School of Medicine and Dentistry, Queen's University Belfast, Belfast, Northern Ireland, United Kingdom. G.Skibinski@qub.ac.uk
Elborn
J Stuart
JS
Ennis
Madeleine
M
D000906:Antibodies / Q000494:pharmacology; D001980:Bronchi / Q000166:cytology* / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects; D006133:Growth Substances / Q000494:pharmacology; D017228:Hepatocyte Growth Factor / Q000378:metabolism*; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology; D018836:Inflammation Mediators / Q000378:metabolism; D009500:Neutralization Tests; D013997:Time Factors
D000906:Antibodies; D017077:Culture Media, Conditioned; D016207:Cytokines; D006133:Growth Substances; D018836:Inflammation Mediators; D017228:Hepatocyte Growth Factor; D015232:Dinoprostone; D007213:Indomethacin