Figure S1: LNCaP, VCaP and DuCaP cells were grown in charcoal-dextran stripped serum (CSS) for 48 h and treated with increasing concentrations of R1881 or DHT in the presence or absence of Enzalutamide (10 µM) or vehicle (Ctrl) for 48 h. Figure S2: Androgens increased lipid uptake of long-chain fatty acids. LAPC4 cells were grown in CSS for 48 h and treated with 1 nM R1881 or vehicle for 48 h. Figure S3: (A) LNCaP cells were synchronized in G0/G1 by androgen deprivation (CSS for 48 h) followed by treatment with Tunicamycin (1 mg/mL), Hydroxyurea (1 M), or Nocodazole (25 ug/mL) for another 24 h, placing cell cycle blocks in G0/G1, S phase and mitosis, respectively. Figure S4: Oncomine analysis of candidate lipid transporters in (A) Grasso [2], (B) Varambally [3] and (C) LaTulippe datasets [4] comparing gene expression of normal prostate gland versus localized, primary prostate cancer tumor samples. Figure S5: (A) DuCaP (top panel) and VCaP cells (bottom panel) were grown in CSS for 48 h and treated with 10 nM DHT in the absence or presence of Enz (10 µM) or vehicle (Ctrl) for 48 h. Table S1. Primer sequences
ARTICLE ABSTRACT
De novo lipogenesis is a well-described androgen receptor (AR)–regulated metabolic pathway that supports prostate cancer tumor growth by providing fuel, membrane material, and steroid hormone precursor. In contrast, our current understanding of lipid supply from uptake of exogenous lipids and its regulation by AR is limited, and exogenous lipids may play a much more significant role in prostate cancer and disease progression than previously thought. By applying advanced automated quantitative fluorescence microscopy, we provide the most comprehensive functional analysis of lipid uptake in cancer cells to date and demonstrate that treatment of AR-positive prostate cancer cell lines with androgens results in significantly increased cellular uptake of fatty acids, cholesterol, and low-density lipoprotein particles. Consistent with a direct, regulatory role of AR in this process, androgen-enhanced lipid uptake can be blocked by the AR-antagonist enzalutamide, but is independent of proliferation and cell-cycle progression. This work for the first time comprehensively delineates the lipid transporter landscape in prostate cancer cell lines and patient samples by analysis of transcriptomics and proteomics data, including the plasma membrane proteome. We show that androgen exposure or deprivation regulates the expression of multiple lipid transporters in prostate cancer cell lines and tumor xenografts and that mRNA and protein expression of lipid transporters is enhanced in bone metastatic disease when compared with primary, localized prostate cancer. Our findings provide a strong rationale to investigate lipid uptake as a therapeutic cotarget in the fight against advanced prostate cancer in combination with inhibitors of lipogenesis to delay disease progression and metastasis.
Prostate cancer exhibits metabolic plasticity in acquiring lipids from uptake and lipogenesis at different disease stages, indicating potential therapeutic benefit by cotargeting lipid supply.