Androgen receptor signalling in macrophages promotes TREM-1-mediated prostate cancer cell line migration and invasion

The androgen receptor (AR) is the master regulator of prostate cancer (PCa) development, and inhibition of AR signalling is the most effective PCa treatment. AR is expressed in PCa cells and also in the PCa-associated stroma, including infiltrating macrophages. Macrophages have a decisive function in PCa initiation and progression, but the role of AR in macrophages remains largely unexplored. Here, we show that AR signalling in the macrophage-like THP-1 cell line supports PCa cell line migration and invasion in culture via increased Triggering Receptor Expressed on Myeloid cells-1 (TREM-1) signalling and expression of its downstream cytokines. Moreover, AR signalling in THP-1 and monocyte-derived macrophages upregulates IL-10 and markers of tissue residency. In conclusion, our data suggest that AR signalling in macrophages may support PCa invasiveness, and blocking this process may constitute one mechanism of anti-androgen therapy.

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ChIP-seq
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Sample preparation Instrument Freshly collected biopsies were chopped in PBS and processed using a gentleMACS Dissociator (Miltenyi Biotec). Programs used for human tissue were "h_tumour_01" for one time and "h_tumour_03" for three times. The cell suspension was subsequently filtered using a 70p.m nylon cell strainer (BD Biosciences) and washed with PBS. Cells were centrifuged at 1200 rpm for 6 min and resuspended in PBS. Cell suspension was stained for flow cytometry sorting with CD14-PE and CD11b-PE antibody (eBioscience) in PBS + 0.5% BSA for 20 min at 4°C LSR Fortessa SORP1 flow cytometer and MoFlo Astrios Beckman Coulter

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Flowlo V9 ieww ns 5uwodaJ LI Die, DSai in 8 1oziaqol po Cell population abundance Abundance of target cells was <1% Gating strategy Macrophage like cells were first selected based on FSC-SSC, followed by doublets exclusion and DAPI-negative selection. CD45cells were then excluded. CD45+ cells with low SCC scatter were excluded as possible T lymphocytes. CD14+ and/or CD11b+ macrophages were isolated in 384 well plates.
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