Project description:We report the genome wide identification of AR binding sites in hormone-deprived A375 melanoma cell line, with either endogenous levels of the lncRNA SLNCR, or a vector expressing SLNCR1.
Project description:Melanoma incidence and mortality rates are historically higher for men than women. Although emerging studies have highlighted tumorigenic roles for the male sex hormone androgen and its receptor (AR) in melanoma, cellular and molecular mechanisms underlying these sex-associated discrepancies are poorly defined. Here, we delineate a previously undisclosed mechanism by which androgen-activated AR transcriptionally upregulates fucosyltransferase 4 (FUT4) expression, which drives melanoma invasiveness by interfering with adherens junctions (AJs). Global phosphoproteomic and fucoproteomic profiling, coupled with in vitro and in vivo functional validation, further reveals that AR-induced FUT4 fucosylates L1 cell adhesion molecule (L1CAM), which is required for FUT4-increased metastatic capacity. Tumor microarray and gene expression analyses demonstrate that AR-FUT4-L1CAM-AJs signaling correlates with pathological staging in melanoma patients. By delineating key androgen-triggered signaling that enhances metastatic aggressiveness, our findings help to explain sex-associated clinical outcome disparities and highlight AR/FUT4 and its effectors as potential prognostic biomarkers and therapeutic targets in melanoma.
Project description:Melanoma incidence and mortality rates are historically higher for men than women. Although emerging studies have highlighted tumorigenic roles for the male sex hormone androgen and its receptor (AR) in melanoma, cellular and molecular mechanisms underlying these sex-associated discrepancies are poorly defined. Here, we delineate a previously undisclosed mechanism by which androgen-activated AR transcriptionally upregulates fucosyltransferase 4 (FUT4) expression, which drives melanoma invasiveness by interfering with adherens junctions (AJs). Global phosphoproteomic and fucoproteomic profiling, coupled with in vitro and in vivo functional validation, further reveals that AR-induced FUT4 fucosylates L1 cell adhesion molecule (L1CAM), which is required for FUT4-increased metastatic capacity. Tumor microarray and gene expression analyses demonstrate that AR-FUT4-L1CAM-AJs signaling correlates with pathological staging in melanoma patients. By delineating key androgen-triggered signaling that enhances metastatic aggressiveness, our findings help to explain sex-associated clinical outcome disparities and highlight AR/FUT4 and its effectors as potential prognostic biomarkers and therapeutic targets in melanoma.
Project description:The text description is: LNCaP are androgen receptor expressing prostate carcinoma cells. Genital skin fibroblasts also express the androgen receptor. Cells were either proliferating or they were G0-arrested. Treatment of cells was performed with either the androgen DHT (dihydrotestosterone), the androgen analogue R1881 (methyltrienolone), or the solvent ethanol. Some hybridizations were performed in the type 1 design. In these cases, the hormone treated sample was hybridized against the ethanol treated sample on the same microarray. Hormone mediated induction or repression of gene transcription can directly be deduced from R/G normalized ratios on arrays. For other experiments, the ethanol treated control and the hormone treated experiment were hybridized on separate microarrays against a common reference of RNA. This reference was composed out of common reference batch CRG (50%) and a fibroblast RNA (50%). This reference was called mixed reference. For all experiments, the same batch of mixed reference was used. A compound treatment design type is where the response to administration of a compound or chemical (including biological compounds such as hormones) is assayed. Compound Based Treatment: Ethanol, R1881 (methyltrienolone), or DHT (dihydrotestosterone) Cell Line: LNCaP (prostate cancer) or foreskin fibroblasts Culture Synchrony: proliferation / Go-arrest Computed
Project description:In this work we reported in a high-throughput way (RIP-seq) the RNAs associated with androgen receptor after treatment with androgen hormone. We used a large compilation of lincRNAs to describe the differences between lincRNAs associated to androgen receptor from those who are non-associated with androgen receptor. By integrating different data sources (DNA-seq Seq and CHIP-seq) was possible to describe transcription factors and histone marks diffrentially enriched at the promoter and vicinity of androgen associated lincRNA loci.
Project description:The text description is: LNCaP are androgen receptor expressing prostate carcinoma cells. Genital skin fibroblasts also express the androgen receptor. Cells were either proliferating or they were G0-arrested. Treatment of cells was performed with either the androgen DHT (dihydrotestosterone), the androgen analogue R1881 (methyltrienolone), or the solvent ethanol. Some hybridizations were performed in the type 1 design. In these cases, the hormone treated sample was hybridized against the ethanol treated sample on the same microarray. Hormone mediated induction or repression of gene transcription can directly be deduced from R/G normalized ratios on arrays. For other experiments, the ethanol treated control and the hormone treated experiment were hybridized on separate microarrays against a common reference of RNA. This reference was composed out of common reference batch CRG (50%) and a fibroblast RNA (50%). This reference was called mixed reference. For all experiments, the same batch of mixed reference was used. A compound treatment design type is where the response to administration of a compound or chemical (including biological compounds such as hormones) is assayed. Compound Based Treatment: Ethanol, R1881 (methyltrienolone), or DHT (dihydrotestosterone) Cell Line: LNCaP (prostate cancer) or foreskin fibroblasts Culture Synchrony: proliferation / Go-arrest Keywords: compound_treatment_design
Project description:We report the dual role of FoxA1 in androgen receptor recruitment to the chromatin of androgen responsive prostate cancer cell line LNCaP-1F5 using ChIP-sequencing. Depletion of FoxA1 reprograms both androgen and glucocorticoid receptor recruitment and subsequent gene expression. The ChIP-seq has been performed using AR, FoxA1, GR, H3K4me2 antibodies. We have also mapped the DNaseI-hypersensitive sites (DHS) using deep sequencing. Examination of AR, FoxA1, GR, H3K4me2 binding sites and DHS sites in parental and FoxA1 depleted LNCaP-1F5 cells.
Project description:Prostate cancer is the most common cancer in men and AR downstream signalings promote prostate cancer cell proliferation. To investigate the AR signaling, we performed ChIP sequence analysis in AR positive prostate cancer cell line, LNCaP. In addition, we used hormone-refractory prostate cancer model cells, Bicalutamide-resistant (BicR) to explore the differences of androgen signaling in prostate cancer progression. ChIP sequence analysis of AR binding sites and epigenetic condition in two prostate cancer cells
Project description:Treatment of late passage (LP50) LNCaP cells with R1881 (androgen) and AR shRNA identified a gene program controlled by androgen receptor in the absence of androgen. Gene expression in late passage (LP50) LNCaP cells that had enhanced androgen-independent growth was determined in androgen-depleted medium in response to R1881 or AR knock down via AR shRNA
Project description:We profiled androgen receptor (AR) genomic targets using high-throughput sequencing of chromatin-immunoprecipitated (ChIP) DNA from TMPRSS2-ERG fusion gene positive DUCaP prostate cancer cells. ChIp-seq and microarray gene expression profiling datasets were integrated with the NHGRI GWAS PCa risk SNPs catalog to identify disease susceptibility SNPs localized within functional androgen receptor binding sites (ARBSs). Eighty GWAS index or linked SNPs were found to be localized in ARBSs. Among these rs11891426:T>G in the 7th intron of the melanophilin gene was found located within a novel putative auxiliary AR binding motif, which we found enriched in the neighborhood of canonical androgen responsive elements. T→G exchange attenuated the transcriptional activity of the ARBS in an AR reporter gene assay of prostate cancer cell models. It went also in line with decreased melanophilin protein level in primary prostate tumors with G allele.These results unravel a hidden link between androgen receptor and a functional PCa risk SNP, whose allele alteration affects androgen regulation of its host gene melanophilin . Genomic profile of androgen receptor binding sites of androgen or vehicle treated DUCaP cells using ChIP-seq. IgG precipiated DNAs from both treatments served as controls.