Project description:Expression data from DHT stimulation vs. control in LNCaP cells LNCaP cells were maintained in phenol red-free RPMI supplemented with 10% charcoal/dextran stripped FCS for three days before stimulation with 100 nM dihydrotestosterone (DHT) for 48 hours
Project description:To analyze the regulatory mechanism of DHT in LNCaP cells (WT) and HOXC9 stably expressed LNCaP cells (C9), we analyzed the effects of 10-7M of DHT on LNCaP cells.
Project description:Microarray experiments were carried out to ascertain whether TOP2β is required for DHT induced androgen receptor target gene expression. We investigated the effect of pharmacological inhibition or RNA interference-mediated depletion of TOP2β on gene expression in androgen-dependent LNCaP prostate cancer cells. Analysis of gene expression in LNCaP cells under various conditions including serum starvation, DHT treatment, and DHT treatment combined with TOPO2B pharmacological inhibitors (Merbarone and Etoposide) and TOPO2B-shRNA knockdown.
Project description:Microarray experiments were carried out to ascertain whether TOP2β is required for DHT induced androgen receptor target gene expression. We investigated the effect of pharmacological inhibition or RNA interference-mediated depletion of TOP2β on gene expression in androgen-dependent LNCaP prostate cancer cells.
Project description:To analyze the regulatory mechanism of DHT and D3 via their respective nuclear receptors in LNCaP cells, we analyzed the effects of those hormones on LNCaP cells when they were treated with siRNA for their respective nuclear receptors.
Project description:Enzalutamide (formerly MDV3100 and available commercially as Xtandi), a novel androgen receptor (AR) signaling inhibitor, blocks the growth of castration-resistant prostate cancer (CRPC) in cellular model systems and was shown in a clinical study to increase survival in patients with metastatic CRPC. Enzalutamide inhibits multiple steps of AR signaling: (1) binding of androgens to AR, (2) AR nuclear translocation, and (3) association of AR with DNA. Here we used Affymetrix human genome microarray technology to investigate the global programme of gene expression of LNCaP cells in response to enzalutamide alone and in the context of DHT-stimulated androgen receptor gene expression. LNCaP cells were grown in RPMI 1640 supplemented with 5% hormone depleted FBS and treated with vehicle (control sample) , DHT (100 nM), enzalutamide (1 or 10 M-BM-5M) or DHT (100 nM) plus enzalutamide (1 or 10 M-BM-5M)for 16 hours for RNA extraction and hybridization. Each condition was done in triplicate.
Project description:ABSTRACT The effects of DHEA were compared with those of DHT using gene expression array profiles in human LNCaP prostate cancer cells. LNCaP cells were exposed to DHEA (300 nM), DHT (300 nM), or vehicle for 48 hrs, and mRNA was isolated. mRNA expression was measured using Affymetrix HU-95 gene chips in 3 experiments performed on different dates. Gene expression values for the two treatment groups and control were sorted in ascending order on the p-values corresponding to a variance stabilized Hotelling test, which measured the extent of differential RNA expression between control and either hormone treatment. The top four genes with significant differential expression were S100 calcium binding protein, neurotensin, 24-dehydrocholesterol reductase, and anterior-gradient 2 homologue. Corresponding per comparison p-values were less than 3 x 10 -5. Nested tests of differential expression between DHEA and DHT treatment revealed significant differences (p < 0.01) for two of the four genes: the S100 calcium binding protein and neurotensin. The microarray findings were confirmed by quantitative RT-PCR. The top 83 genes found to exhibit differential expression were used in a pathway analysis. In general, DHT decreased expression of more genes involved in intercellular communication, signal transduction, nucleic acid binding and transport, and in structural components, such as myosin and golgin, than did DHEA. These data reveal consistent, measurable differences in gene expression patterns following treatment of LNCaP prostate cancer cells with DHEA versus DHT. The mechanisms underlying these observations, and the possible pathophysiological significance of these differences, remain to be determined. Keywords: differential gene expression between DHT and DHEA
Project description:Enzalutamide (formerly MDV3100 and available commercially as Xtandi), a novel androgen receptor (AR) signaling inhibitor, blocks the growth of castration-resistant prostate cancer (CRPC) in cellular model systems and was shown in a clinical study to increase survival in patients with metastatic CRPC. Enzalutamide inhibits multiple steps of AR signaling: (1) binding of androgens to AR, (2) AR nuclear translocation, and (3) association of AR with DNA. Here we used Affymetrix human genome microarray technology to investigate the global programme of gene expression of LNCaP cells in response to enzalutamide alone and in the context of DHT-stimulated androgen receptor gene expression.