Receptor-mediated and protein kinase-dependent growth enhancement of primary human fibroblasts by platelet activating factor

✍ Scribed by Steffany A. L. Bennett; H. Chaim Birnboim

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 186 KB
• Volume: 20
• Category: Article
• ISSN: 0899-1987
• DOI: 10.1002/(sici)1098-2744(199712)20:4<366::aid-mc6>3.0.co;2-h

✦ Synopsis

Chronic inflammation is a recognized risk factor for human cancer, but the causal mechanisms are poorly understood. We previously demonstrated that platelet activating factor (PAF) can induce alterations in the in vitro growth properties of primary rat fibroblasts. In the study reported here, exposure of primary human skin fibroblasts to PAF for 1 h in serum-free medium was shown to cause sustained proliferation over 50 d in medium containing low serum and anchorage-independent growth in soft agarose. Both properties could be inhibited by pretreatment with a PAF receptor antagonist, CV3988 (10 µM); a tyrosine-kinase inhibitor, genistein (1 µg/mL); or a protein kinase C (PKC) inhibitor, staurosporine (50 nM) but not with a cyclooxygenase inhibitor, indomethacin (200 nM-20 µM). PAF had no effect on doubling time, saturation density, or cell viability under normal monolayer growth conditions in complete medium. Treatment with lyso-PAF, an inactive metabolite of PAF, had no effect in either of the assays. Control and PAF-induced cell proliferation in low-serum medium was inhibited by PAF receptor antagonists present during the extended growth period. The presence of PAF receptor mRNA in human skin fibroblasts was demonstrated by reverse transcriptase-polymerase chain reaction. The presence of a functional receptor was indicated by an early (2 min) transient increase in PKC activity and an increase in fos mRNA after PAF treatment. PAF-induced PKC activity was blocked by pretreatment with either staurosporine (50 nM) or CV3988 (1 µM). These results suggest that PAF is a mitogenic factor that contributes to the known increase in risk of malignancy associated with chronic inflammatory conditions.