male prostate cancer cell line vcap  (ATCC)

Journal: Cancer research

Article Title: Androgen receptor drives polyamine synthesis creating a vulnerability for prostate cancer

doi: 10.1158/0008-5472.CAN-25-1532

Figure Lengend Snippet: A. Tumor size over time of SKCaP-1R patient-derived xenograft tumors growing untreated (Control) or treated with supraphysiological androgen (SPA; testosterone cypionate subcutaneous pellet). B. Principal component analysis with 95% confidence ellipses of metabolite abundance in tumors from (A). C. Volcano plot displaying change in metabolite abundance in SKCaP-1R tumors treated with SPA versus Control from (A). D. Polyamine synthesis pathway schematic displaying metabolites altered in abundance by SPA in tumors from (A). E. Isotope tracing schematic. Cells were treated with either vehicle control (VEH; EtOH 0.1%) or SPA (R1881 10nM) in media containing either uniformly-labeled 13 C-arginine or 13 C-putrescine Metabolites were extracted at 24 hours. F-G. Contribution of polyamine synthesis pathway metabolites from arginine (F) or putrescine (G) in LNCaP and VCaP cells treated with VEH or SPA as per experimental design of (E). N=3 independent experiments. P values by unpaired 2-tailed t test.

Article Snippet: LNCaP and VCaP cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Derivative Assay, Control, Labeling

Journal: Cancer research

Article Title: Androgen receptor drives polyamine synthesis creating a vulnerability for prostate cancer

doi: 10.1158/0008-5472.CAN-25-1532

Figure Lengend Snippet: A. The top 3 tracks display AR ChIP-seq of VCaP cells treated with vehicle control, supraphysiological androgen (SPA; DHT 10nM), or SPA and enzalutamide (ENZA; 10uM) for 12 hours. Data reanalyzed from Asangani et al, 2014( 35 ). Lower tracks display AR ChIP-seq of 7 histologically normal prostate samples and 13 local prostate cancer samples. Data reanalyzed from Pomerantz et al, 2015( 36 ). B. Schematic depicting dCas9-KRAB-mediated blockade of AR binding. C. Protein expression of ODC by western blot of LNCaP cells expressing dCas9-KRAB and indicated sgRNA treated with vehicle control (V) or supraphysiological androgen (S; R1881 10nM) for 24 hours. Vinculin is used as a loading control. Total ODC quantification and ODC fold-change (FC) by SPA of n = 3 independent experiments are displayed below with p values by unpaired 2-tailed t test.

Article Snippet: LNCaP and VCaP cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Binding Assay, ChIP-sequencing, Control, Expressing, Western Blot

Journal: Cancer research

Article Title: Androgen receptor drives polyamine synthesis creating a vulnerability for prostate cancer

doi: 10.1158/0008-5472.CAN-25-1532

Figure Lengend Snippet: A. MYC ChIP-seq in VCaP cells treated with vehicle control or supraphysiological androgen (SPA; DHT 10nM) for 4 hours. Data reanalyzed from Guo et al, 2021( 40 ). B. Change in indicated transcript expression by RNAseq in VCaP cells with control knock-down or MYC knock-down by siRNA. Data reanalyzed from Guo et al, 2021( 40 ). C . Expression of indicated proteins in LNCaP cells expressing empty vector (EV) or MYC (MYC) treated with vehicle control (V; EtOH 0.1%) or SPA (S; R1881 10nM) for 96 hours. Vinculin is used as a loading control. Representative blot of n=2 experiments. D. Change in indicated transcript expression by RNAseq in cells treated as per C. E. A comparison of change of individual RNA transcript abundance by SPA in LNCaP-EV cells versus LNCaP-MYC cells. The line of best fit (dotted red) has a shallower slope than the line of unity (solid blue), indicating that many transcripts have reduced change by SPA in LNCaP-MYC cells. F. Schematic highlighting that the inhibition of MYC by SPA can function as an amplifying circuit to further increase expression of ODC1 , AMD1 , and KLK3 by SPA. P values by unpaired 2-tailed t test in B and D.

Article Snippet: LNCaP and VCaP cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, ChIP-sequencing, Control, Knockdown, Plasmid Preparation, Comparison, Inhibition

Journal: Cancer research

Article Title: Androgen receptor drives polyamine synthesis creating a vulnerability for prostate cancer

doi: 10.1158/0008-5472.CAN-25-1532

Figure Lengend Snippet: A . Principal component analysis of RNAseq of LNCaP cells treated with vehicle control (VEH; EtOH 0.1%), supraphysiological androgen (SPA, R1881 10nM), DFMO (5mM), DFMO and putrescine (PUT; 100uM), DFMO and SPA (SD), and DFMO and SPA and PUT for 96 hours. B-C. Volcano plot displaying change in transcript abundance in LNCaP cells treated with DFMO and SPA (SD) versus VEH (B) or versus SPA (C) as per A. D. RNA and protein expression of MYC of LNCaP and VCaP cells with indicated treatments for 96 hours. MYC RNA expression was normalized to ACTB and MYC protein expression was normalized to vinculin. P values by unpaired 2-tailed t test. E. Volcano plot displaying normalized enrichment scores of HALLMARK gene sets in LNCaP cells treated with DFMO and SPA versus VEH as per (A). F. Abundance of proteins with monomethyl arginine and H4-SDMA in LNCaP and VCaP cells with indicated treatments for 96 hours. G . Protein expression of MYC and AMD1 by western blot in LNCaP and VCaP cells treated with indicated treatments, including spermidine (SPD; 100uM), spermine (SPM; 100uM), toluenesulfonic acid (TSA; 200uM), S-adenosyl-methionine-toluenesulfonic acid (TSA-SAM; 25uM, 50uM, 100uM, 200uM), SAM486 (2uM), and aminoguanidine (1mM) as indicated, for 96 hours. H. Schematic displaying effects of SPA and DFMO combination treatment.. Vinculin is used as a loading control in F and H, where blots are representative of n = 2 or 3 experiments.

Article Snippet: LNCaP and VCaP cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Inhibition, Control, Expressing, RNA Expression, Western Blot