Androgen refractory cancer of the prostate (ARCaP) cells contain androgen receptor (AR) and synthesize and secrete prostate specific antigen (PSA). We isolated epithelia-like ARCaPE from parental ARCaP cells and induced them to undergo epithelial–mesenchymal transition (EMT) by exposing these cells to soluble factors including TGFβ1 plus EGF, IGF-1, β2-microglobulin (β2-m), or a bone microenvironment. The molecular and behavioral characteristics of the resultant ARCaPM were characterized extensively in comparison to the parental ARCaPE cells. In addition to expressing mesenchymal biomarkers, ARCaPM gained 100% incidence of bone metastasis. ARCaPM cells express receptor activator of NF-κB ligand (RANKL), which was shown to increase tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts in culture, and when metastatic to bone in vivo. We provide evidence that RANKL expression was promoted by increased cell signaling mediated by the activation of Stat3-Snail-LIV-1. RANKL expressed by ARCaPM cells is functional both in vitro and in vivo. The lesson we learned from the ARCaP model of EMT is that activation of a specific cell signaling pathway by soluble factors can lead to increased bone turnover, mediated by enhanced RANKL expression by tumor cells, which is implicated in the high incidence of prostate cancer bone colonization. The AR-CaP EMT model is highly attractive for developing new therapeutic agents to treat prostate cancer bone metastasis.
Keywords: EMT, Cell signaling, RANKL, Osteoclastogenesis, LIV-1, Bone metastasis, Prostate, Breast, Lung, Kidney, TGFβ1, EGF