EFFECTS OF GENISTEIN, TYRPHOSTIN, AND PERTUSSIS TOXIN ON EGF-INDUCED MITOGENESIS IN PRIMARY CULTURE AND CLONAL OSTEOBLASTIC CELLS

Epidermal growth factor (EGF) has been found to stimulate proliferatio n in a variety of cell types. The EGF receptor is known to have tyrosi ne kinase activity [1], however, the role of this signal mechanism has not been established in bone cells. The aim of this study was to dete rmine whether tyrosine kinase activity and G inhibitory (Gi()) protein s are involved in EGF-stimulated proliferation in the osteoblastic cel l line G292 and in primary culture osteoblasts isolated from neonatal rat calvaria. Cell proliferation was measured by H-3-thymidine incorpo ration using liquid scintillation spectrometry. EGF stimulates a dose- dependent increase in proliferation of G292 and primary culture cells above control. Genistein was able to inhibit the effects of EGF in the G292 cells. In the primary culture cells, genistein with EGF appeared to enhance proliferation compared with EGF alone or genistein alone. Tyrphostin 25, on the other hand, inhibited the EGF response in both o f these cell types. Inactivation of G(i) proteins with pertussis toxin was able to inhibit EGF-induced mitogenesis in the neonatal rat osteo blasts but did not appear to specifically inhibit this response in the G292 cells. These results suggest that although bath of these osteobl astic cell types increase proliferation in response to EGF, their sign al pathways are different.