Background: The underlying mechanisms permitting prostate cancer bone metastasis are poorly understood. We previously showed that the highly metastatic prostate cancer cell line, PC-3, inhibits bone marrow endothelial (HBME-1) cell growth in collagen gels and induces them to differentiate into cords, resembling angiogenesis in vivo.
Methods: cDNA microarray analysis was performed to identify cytokines responsible for the effects of PC-3 cells on HBME-1 cells. Cytokine and neutralizing antibody studies were done to further investigate specific angiogenic factors, such as transforming growth factor beta (TGFbeta). TGFbeta RNA and protein were detected by real-time RT-PCR and enzyme-linked immunosorbent assay (ELISA) analysis to measure their production by prostate cancer cell lines. Conditioned media experiments using TGFbeta neutralizing antibodies were used to analyze TGFbeta activation by prostate cancer cells.
Results: PC-3 conditioned media altered the expression of several TGFbeta-regulated or -associated genes in HBME-1 cells. Low concentrations of TGFbeta cytokines inhibited HBME-1 cell growth to a similar level as PC-3 conditioned media and partially induced differentiation. Inhibitors and neutralizing antibodies directed against TGFbeta isoforms and TGFbeta receptor type 2 (TGFbetaRII) reversed the growth inhibition of HBME-1 cells conferred by PC-3 conditioned media. Yet, only TGFbetaRII neutralizing antibodies significantly inhibited HBME-1 differentiation. Also, prostate cancer cell lines produced low levels of TGFbeta RNA and protein, and were shown to activate serum-derived TGFbeta.
Conclusions: These results suggest that prostate cancer cells mediate growth inhibition and differentiation of bone marrow endothelial cells both through production and activation of TGFbeta as well as alteration of TGFbetaRII-mediated signal transduction. This could contribute to the establishment and growth of bone metastases.
Prostate 66:632-650, 2006. (c) 2005 Wiley-Liss, Inc.