Project description:The androgen receptor is considered as the key promoter of prostate cancer. It is a transcription factor that controls the transcription of hundreds of its target genes. In this project we focuses on how androgen receptor stimulation by the synthetic androgen R1881 can affect the proteome of peroxiosmes and the antioxidant enzymes in LNCaP cells.
Project description:Time series of the prostate cancer cell line LNCaP, treated for 2, 4, 6 and 8 hours with the synthetic androgen R1881. As control, the cells were cultured for 2, 4, 6 and 8 hours in the presence of the same concentration of solvent (ethanol). With this short treatment time, we aimed to identify mainly direct targets of the androgen receptor. LNCaP has a very low growth rate in steroid stripped medium and resumes growth on addition of androgens. Keywords: Time course
Project description:Androgens are required for the development of normal prostate, and they are also linked to the development of prostate cancer. We used microarrays to understand the role of androgen in an androgen dependent, androgen receptor (AR) positive human metastatic cell line, LNCaP. LNCaP cells were grown in RPMI medium and they were subjected to stay in phenol-red free, RPMI with charcoal stripped serum for 48h. Synthetic androgen R1881 was added and the cells were allowed to grow for 48h. Control cells were given with corresponding amount of ethanol as vehicle which is used for the solubilization of R1881. Cells were harvested and RNA was isolated for microarray analysis.
Project description:LNCaP prostate cancer cells were stimulated with the synthetic androgen R1881. RNA-sequencing was performed to identify changes induced by enzalutamide treatment on the transcriptome level.
Project description:Detailed analysis of androgen regulated gene expression in the LNCaP prostate cancer cell line. Since androgens and the AR are known to be important for prostate cancer cell proliferation and invasion we aimed to identify androgen receptor (AR) regulated genes by combining this detailed Illumina beadarray study of androgen regulated gene expression with AR ChIP-sequencing data. LNCaP cells were grown in RPMI medium supplemented with 10% charcoal dextran stripped (steroid depleted) FBS for 72h, before treatment with 1nM R1881 or vehicle control. Total RNA was harvested every 30min for 4h and then every hour up to 24h using trizol (Sigma), quantified using a Nanodrop spectrophotometer (ND-1000). RNA samples were prepared for analysis on Illumina Human 6 v2 BeadArrays and Biotrove Realtime PCR panels according to the manufacturers protocols (see Supplementary Methods for details). Expression array data were normalised in R using the beadarray software and BASH (see Supplementary Methods for details) (Cairns et al., 2008; Dunning et al., 2007).
Project description:RNA-seq data were obtained from hTERT immortalized human prostate transit amplifying EP156T cells (+/- 10 nM R1881 for 48 hrs), progeny tumorigenic EPT3-M1 cells recovered from mouse metastatic tumor (+/- 10 nM R1881 for 48 hrs) and the prostate cancer cell lines LNCaP (+/- 10 nM R1881 for 48 hrs), VCaP (+/- 1 nM R1881 for 24 hrs) and 22Rv1 (+/- 1 nM R1881 for 24 hrs) (obtained from the American Type Culture Collection). All cells were either stimulated or not stimulated with the synthetic androgen R1881 (1 or 10 nM for 24 or 48 hrs) prior to lysis (Qiagen miRNeasy minikit lysis buffer) and total RNA purification and DNase treatment.
Project description:Transcriptional profiling of LNCaP prostasphere-forming cells, comparing control untreated sphere-forming cells with hormone treated sphere-forming cells. Both estrogen (estradiol) and androgen (R1881) promote the sphere formation of human prostate cancer cell LNCaP. Goal was to determine the effects of hormones on global gene expression of LNCaP sphere-forming cells.
Project description:Transcriptional profiling of LNCaP prostasphere-forming cells, comparing control untreated sphere-forming cells with hormone treated sphere-forming cells. Both estrogen (estradiol) and androgen (R1881) promote the sphere formation of human prostate cancer cell LNCaP. Goal was to determine the effects of hormones on global gene expression of LNCaP sphere-forming cells. Five samples were analyzed. Control (ethanol 1hr), estradiol 1 hour, estradiol 24 hour, R1881 1 hour, R1881 24 hour.
Project description:Analysis of LNCaP cell molecular differences and their response to R1881 in 2D and 3D cultures. Androgen regulated genes were differentially expressed between 2D and 3D cultures. These results provide insights into factors that influence the expression of androgen regulated genes In this study, LNCaP cells cultured in tissue culture plastic (2D) and in the hydrogel (3D) were maintained up to 3 days and 24 days respectively in serum containing media before they were androgen-starved for 48 hours. Cells were either treated with 1nM R1881 or continued to be androgen-deprived (without R1881 with 0.008% ethanol) for another 48 hours prior to cell harvest for gene expression analysis. Biological triplicates were prepared for each condition.