Project description:Axl expression is deregulated in several cancer types, predicts poor overall patient survival and is linked to increase in resistance to drug therapy. Here, we have evaluated a library of natural compounds for inhibitors of Axl. We identified dihydroartemisin (DHA), the active principle of the anti-malarial drug artemisinin, as a specific Axl inhibitor in prostate cancer (PCa). DHA blocks Axl expression leading to apoptosis induction, decrease in cell proliferation, migration and tumor development of PCa cells. Furthermore, DHA treatment synergizes with docetaxel, a standard of care in metastatic PCa, and increases the overall survival of mice with human PCa xenografts. We demonstrated that DHA-mediated control of miR-34a and miR-7 expression leads to inhibition of Axl expression. This process is dependent on regulation of the chromatin via methylation of histone H3 lysine 27 residues (H3K27) by Jumonji, AT-rich interaction domain containing 2 (JARID2) and the enhancer of zeste homolog 2 (EZH2), components of the Polycomb Complex Repressor 2 (PCR2). Our discovery of a previously unidentified miR-34a/miR-7/JARID2 pathway that controls DHA-mediated effects on Axl expression and inhibition of cancer cell proliferation, migration, invasion and tumor formation provides new molecular and mechanistic insights into DHA’s anti-cancer effect on PCa with potential therapeutic implications.
Project description:To investigate the anti-tumor function of compound H93 in the development of prostate cancer, we treated DU145 cell lines with H93, SAHA or DMSO. We then performed gene expression profiling analysis using data obtained from RNA-seq of cells after treatment.
Project description:To investigate the anti-tumor function of compound H93 in the development of prostate cancer, we treated DU145 cell lines with H93, SAHA or DMSO. We then performed genome wide DNA methylation profiling analysis of cells after treatment. The Illumina Infinium MethylationEPIC BeadChip (850K BeadChip) was used to obtain DNA methylation profiles across approximately 850,000 CpGs.
Project description:To uncover the mechanism underlying the effect of pentamidine on prostate cancer cells, we performed RNA sequencing (RNA-seq) to compare the transcriptional difference between pentamidine- and vehicle-treated prostate cancer cells (PC3 and DU145 cells).
Project description:DU145 prostate cancer cells were treated with 25 ng/ml hepatocyte growth factor (HGF) or vehicle for 2, 8, or 24 hours. HGF stimulates the cMET protein, a tyrosine kinase transmembrane protein. The aim of this study is to determine the role of the HGF/cMET pathway in immature cells of established prostate cancer. HGF stimulation of DU145 prostate cancer cell line led to cell migration in culture, formation of sprouts in Matrigel and inhibition of growth. These biological effects went together with induction of a stem-like phenotype as defined by up-regulation of CD49b, CD49f, CD44 and SOX9, and down-regulation of CD24 on gene-expression arrays and quantitative PCR. The shift towards a stem-like phenotype was reflected by protein modifications on FACS, Western blot, and enhanced rapid adhesion to collagen I. Small molecules SU11274 and PHA665752 were able to inhibit both morphologic and molecular HGF effects.
Project description:DU145 prostate cancer cells were treated with 25 ng/ml hepatocyte growth factor (HGF) or vehicle for 2, 8, or 24 hours. HGF stimulates the cMET protein, a tyrosine kinase transmembrane protein. The aim of this study is to determine the role of the HGF/cMET pathway in immature cells of established prostate cancer. HGF stimulation of DU145 prostate cancer cell line led to cell migration in culture, formation of sprouts in Matrigel and inhibition of growth. These biological effects went together with induction of a stem-like phenotype as defined by up-regulation of CD49b, CD49f, CD44 and SOX9, and down-regulation of CD24 on gene-expression arrays and quantitative PCR. The shift towards a stem-like phenotype was reflected by protein modifications on FACS, Western blot, and enhanced rapid adhesion to collagen I. Small molecules SU11274 and PHA665752 were able to inhibit both morphologic and molecular HGF effects. DU145 cells were stimulated for 2, 8 and 24 hours with 25 ng/ml HGF or vehicle. For each time point two arrays analyses were performed. One for cells stimulated with a vehicle and one for the HGF stimulated cells. Six arrays were performed in total in this study.
Project description:Activation of Signal Transducer and Activator of Transcription 3 (STAT3) is common in prostate cancers. STAT3 may induce cell proliferation and resistance to apoptosis, as well as promote tumor angiogenesis, invasion, and migration by activating gene expression. Many STAT3-dependent transcriptional responses are mediated through protein-protein interactions that involve the amino-terminal domain (N-domain). In this study, we found that inhibition of the STAT3 N-domain using novel inhibitor ST3-Hel2A-2 induces apoptotic death in prostate cancer cells. The cell death was accomponied by robust activation of pro-apoptotic gene. Using chromatin immunoprecipitation and tiling human promoter arrays (ChIP-chip), we have defined genome-wide targets of STAT3 in DU145 prostate cancer cells. We found that upregulated pro-apoptotic genes were bound by STAT3 in prostate cancer cells, and that STAT3 binding was decreased following inhibition of the STAT3 N-domain. STAT3 siRNA knockdow confirmed specificity of STAT3 binding and changes in gene expression. DU145 cells were treated with STAT3 siRNA or scrambled siRNA for 48hr. Total RNA has been extracted and prepared for hybridization on Affymetrix HG-U133A 2.0 arrays.
Project description:Prostate cancer stem-like cells were derived from DU145 cells as suspension spheres. DU145 cells were transduced with a CNTN1 over expressing retroviral vector and a control empty vector. DU145 spheres were transduced using a retroviral-based shRNA vector against CNTN1 and a scrambled control shRNA viral vector. Both cell lines were cultured over a couple of passages before RNA was collected.
Project description:The role of a number of miRNAs found to be differentially expressed in prostate cancer was functionally investigated by overexpressing them in DU145 prostate cancer cells and by analyzing gene expression profiles.