PDF file, 1659K, S1. Effects of THP-1 on cell migration, acinar morphogenesis, and downstream signaling in BPH-1 cells. S2. H&E staining of the anterior prostate from athymic nude mice 60 days after orthoptoic injection of either THP-1 or RWPE-1 cells. S3. Elevated pSTAT3 in mouse prostate epithelial cells (mPrE) after co-culture with mouse macrophages. S4. THP-1 cells were cultured alone for 48 hrs, and the expressions of cytokines were determined by a human cytokine array (R&D Systems). S5. qPCR (A) and Western blot analysis (B) of p53 and PTEN expression levels in RWPE-1 cells and RWPE-1/THP-1 cells after long term co-culture for 5 days. S6. The role of CCL4 and macrophage AR in the modulation of EMT markers and CCR1 expression in RWPE-1 cells. S7. Effects of macrophages AR silencing on BPH-1 cells during co-culture. S8. Generation and confirmation of MARKO/PTEN+/- mice. S9. CCL4 expression in prostate cancer does not associate with pathologic stage, lymph node metastasis, and recurrent-free survival. S10. IHC Staining of Mouse PTEN knockout prostate using an anti-Snail antibody.
ARTICLE ABSTRACT
Infiltrating macrophages are a key component of inflammation during tumorigenesis, but the direct evidence of such linkage remains unclear. We report here that persistent coculturing of immortalized prostate epithelial cells with macrophages, without adding any carcinogens, induces prostate tumorigenesis and that induction involves the alteration of signaling of macrophage androgen receptor (AR)-inflammatory chemokine CCL4–STAT3 activation as well as epithelial-to-mesenchymal transition and downregulation of p53/PTEN tumor suppressors. In vivo studies further showed that PTEN+/− mice lacking macrophage AR developed far fewer prostatic intraepithelial neoplasia (PIN) lesions, supporting an in vivo role for macrophage AR during prostate tumorigenesis. CCL4-neutralizing antibody effectively blocked macrophage-induced prostate tumorigenic signaling and targeting AR via an AR-degradation enhancer, ASC-J9, reduced CCL4 expression, and xenografted tumor growth in vivo. Importantly, CCL4 upregulation was associated with increased Snail expression and downregulation of p53/PTEN in high-grade PIN and prostate cancer. Together, our results identify the AR-CCL4-STAT3 axis as key regulators during prostate tumor initiation and highlight the important roles of infiltrating macrophages and inflammatory cytokines for the prostate tumorigenesis. Cancer Res; 73(18); 5633–46. ©2013 AACR.
