ABSTRACT: RD induced DNA damage and suppressed DNA repair in PC-3 cells. Expression profiling of PC-3 cells treated with 10 umol/L RD for 24 hours. Total RNA was extracted from PC-3 cells exposed to RD for 24 h using an RNAiso plus kit (TaKaRa). Microarray analysis was performed on separate samples on the Affymetrix HG-U133 plus 2 chip.
Project description:RD induced DNA damage and suppressed DNA repair in PC-3 cells. Expression profiling of PC-3 cells treated with 10 umol/L RD for 24 hours. Overall design: Total RNA was extracted from PC-3 cells exposed to RD for 24 h using an RNAiso plus kit (TaKaRa). Microarray analysis was performed on separate samples on the Affymetrix HG-U133 plus 2 chip.
Project description:Metastatic prostate cancer is a leading cause of cancer-related death in men. Cancer stem cells (CSCs) are involved in tumor progression and metastasis, including in prostate cancer. There is an obvious and urgent need for effective cancer stem cells specific therapies in metastatic prostate cancer. MicroRNAs (miRNAs) are an important class of pervasive genes that are involved in a variety of biological functions, especially in cancer. The goal of this study was to identify miRNAs involved in prostate cancer metastasis and cancer stem cells. Several published reports have demonstrated that non-adherent spheres culture is increasingly used as an effective method to enrich and identify stem cells or putative CSCs.In our previous study, we enriched prostate cancer stem cells from PC-3 sphere cells in serum-free suspension culture and characterized their CSCs properties.Thus, we used spheres as a prostate cancer stem cells model to elucidate its metastatic mechanisms. We examined the miRNA expression profiles of PC-3 sphere cells of prostate cancer compared with PC-3 adherent cells by miRNA microarray.
Project description:To determine if any altered miRNA expression is actually involved in the growth inhibition and/or cell death induction by luteolin and/or gefitinib, we performed miR-array analysis using RNA from PC-3 cells after 24h of treatment with 60μM luteolin and/or 60μM gefitinib. PC-3 cells were treated for 24h with DMSO, 60μM luteolin (Lut), 60μM gefitinib (Gef) or their co-administration (Lut+Gef). Each sample was run in duplicate.
Project description:Transcriptional profiling of the prostate adenocarcinoma cell line PC-3 comparing control cells transfected with a pool of oligonucleotides control (siControl) with cells transfected a pool of oligonucleotides specific to inhibit ARHGAP21 expression (siARHGAP21). The efficiency of the transfection was checked with quantitative PCR and western blotting. Goal was to determine the effects of ARHGAP21 silencing on global PC-3 gene expression. Two-condition experiment, siControl vs. siARHGAP21 PC-3 cells with three biological samples of PC3 cells transfected with a pool of oligonucleotides control (siControl 1, 2 and 3) and three biological samples of PC3 cells transfected with a pool of oligonucleotides specific to inhibit ARHGAP21 (siARHGAP21 1, 2 and 3). For each sample (three siControl and three siARHGAP21), two technical replicates were performed (labelled with Cy3 or Cy5).
Project description:Transforming growth factor-β (TGF-β) is a key factor for the development of prostate cancer metastases in bone. In breast cancer and melanoma, studies have shown how TGF-β regulates gene expression to allow cancer cells to adapt to the bone microenvironment. We used microarray analyses to characterize the effect of TGF-β on gene expression in prostate cancer cells in vitro. Human prostate cancer cells, PC-3, were cultured in the presence or absence of TGF-β. Total RNA was extracted for hybridization on Affymetrix microarray analyzed with Microarray Analysis Suite 5.0.
Project description:Genome wide expression changes following 50uM Casodex treatment were investigated to determine regulatory targets commonly overlooked in gene function specific microarrays and for comparison of the effects the wild-type AR expressing PC-346C cells against the mutant androgen receptor (AR) LNCaP cell line. PC-346C Prostate Cancer cells were treated for a period of 48h with or without 50uM Casodex following a 36h seeding period. At the selected time point, total RNA was harvested from the cells for hybridization and analysis by Nimblgen Systems Inc using the homo sapiens gene expression array.
Project description:Prostate cancer is the leading type of cancer diagnosed and the third leading cause of cancer-related deaths worldwide each year in men. The limitations of the current prostate cancer screening test demands new biomarkers for early diagnosis of prostate cancer metastasis to bone. In this study, we performed a deep proteomic analysis of secreted proteins from the prostate cancer bone metastasis cell line, PC-3, and normal prostate cell line, RWPE-1. Here, we quantified 917 proteins and found 68 highly secreted in PC-3 versus RWPE-1 cells using LC-MS/MS. To characterize the highly secreted proteins in the PC-3 cell line to identify biomarker proteins, the quantifiable proteins were divided into four quantitative categories (Q1-Q4). The KEGG pathways of lysine degradation and osteoclast differentiation were enriched in Q4, the highly secreted group. Transforming growth factor (TGF) beta family proteins related to osteoclast differentiation were identified as key regulators in PC-3 cells. Among the 68 highly secreted proteins, pentraxin, follistatin, and TGF-beta family members were confirmed by immunoblots. In particular, serpin B3, modulated by TGF-beta, was detected and its selective expression and secretion in PC-3 cells was confirmed. In the present study, we suggest that serpin B3 is a novel biomarker candidate for diagnosis of prostate cancer metastasis to the bone.
Project description:The prostate cancer cell line PC-3 was transfected with pre-miR-375 or control. After 48 hours the cells were lysed and immunoprecipitation was performed using anti-panAgo (...) or isotype anti-IgG antibodies overnight. RNA of total lysates and immunoprecipitation fractions was extracted using miRNeasy kit (Qiagen) and libraries were generated using the Stranded Total RNA Sample Prep Kit (Takara Clontech). The experiment was performed in order to identify potential targets of miR-375 in prostate cancer.
Project description:To determine the global effects of ASCT2 inhibition, we used next generation sequencing to determine mRNA expression changes in PC-3 cells treated with BenSer or GPNA for 48 h. Examination of two different ASCT2 inhibitors BenSer and GPNA in prostate cancer cell line PC-3.