Increased PTHRP production by a tyrosine kinase oncogene, Tpr-Met: role of the Ras signaling pathway.

We have used the Tpr-Met oncogene as a model to examine signaling pathways of growth factors and tyrosine kinase oncogenes that can increase parathyroid hormone-related peptide (PTHRP) production. PTHRP production in Tpr-Met transfected cells, when assessed by Northern blot analysis and radioimmunoassay, was increased four- to eightfold. Treatment of these cells with the transcriptional inhibitor actinomycin D and nuclear run-off assays showed that the major cause of increased PTHRP mRNA was enhanced gene transcription. To analyze the intracellular signaling molecules involved in PTHRP production, stable cell lines expressing a Tyr489 Phe mutant of the Tpr-Met oncoprotein were examined. The mutant fails to activate phosphatidylinositol (PI)-3 kinase or associate with the Grb-2 adaptor protein and caused a significant reduction in PTHRP production. Treatment of wild-type Tpr-Met transfected cells with wortmannin, a PI-3 kinase inhibitor, had no effect on PTHRP production; however, treatment of these cells with lovastatin, an inhibitor of p21ran isoprenylation, significantly reduced PTHRP expression. These results show that PTHRP is a downsteam target of the Tpr-Met oncogene and indicate that the PTHRP stimulating activity is mediated via the Ras signaling pathway.