Project description:To identify genes regulated by GRK3 in human cancer cells. Extracted total RNAs from cultured cells, processed samples and hybridized on Illumina Ref8-v3 BeadChips
Project description:The biguanide metformin has been shown to not only reduce circulating glucose levels but also suppress in vitro and in vivo growth of prostate cancer. However, the mechanisms underlying the anti-tumor effects of metformin in advanced prostate cancers are not fully understood. The goal of the present study was to define the signaling pathways regulated by metformin in androgen-receptor (AR) positive, castration-resistant prostate cancers. Our group used RNA sequencing (RNA-seq) to examine genes regulated by metformin within the C4-2 human prostate cancer cell line. Western blot analysis and quantitative RT-PCR were used to confirm alterations in gene expression and further explore regulation of protein expression by metformin. Data from the RNA-seq analysis revealed that metformin alters the expression of genes products involved in metabolic pathways, the spliceosome, RNA transport, and protein processing within the endoplasmic reticulum. Gene products involved in ErbB, insulin, mTOR, TGF-, MAPK, and Wnt signaling pathways are also regulated by metformin. A subset of metformin-regulated gene products were genes known to be direct transcriptional targets of p53 or AR. Together, our results suggest metformin regulates multiple pathways linked to tumor growth and progression within advanced prostate cancer cells.
Project description:Prostate cancer (PCa) is the most common cancer in American men. The American Cancer Society’s estimates for prostate cancer in the United States for 2017 are estimated 161.360 new cases and 26,730 deaths from PCa. To study metastatic properties to bone, PC-3 cell line is mainly used classical human prostatic carcinoma cell line, established and characterized its tumorigenicity from a human prostatic adenocarcinoma metastatic to bone is reported. In addition, PC-3/nkR cell line, natural killer(NK) cells-resistant, was isolated from mammary tumor xenograft studies in mice from PC-3 was implanted to nude mice and fecund to be tumorigenic in the early 2000s. In this study, we investigated secreted proteins of the conditioned media of PC-3 and PC-3/nkR cell lines using comparative proteomics technology to identify the molecular mechanism related to metastatic processes related to PC-3/nkR. Our study showed PC-3/nkR cells are new highly migrated and NK cells-resistant cell-line compared to PC-3 cells, as novel highly malignant tumor cells to study mechanisms of PCa metastatic.
Project description:The aberrant activation of the ERG oncogenic pathway due to TMPRSS2-ERG gene fusions is the major driver of prostate cancer initiation and progression. We identified the alpha1 and beta1 subunits of soluble guanylyl cyclase (GUCY1A1, GUCY1B1) as major ERG-regulated genes in prostate cancer cells. Soluble guanylyl cyclase (sGC) is the major mediator of nitric oxide signaling in cells that, upon nitric oxide binding, catalyzes the synthesis of cGMP and subsequently activates PKG. We showed in ERG-positive PCa cells (VCaP) that cGMP synthesis was significantly elevated by ERG, leading to increased PKG activity and cell proliferation. To further understand the functions of sGC-cGMP pathway in prostate cancer cells, we performed RNA-seq analyses in VCaP cells to identify genes that are regulated by sGC.
Project description:The objective of this study was to evaluate the consequences of Ago1 knockdown in metastatic prostate cancer cells PC3. To this end we profiled RNA from PC3 cells 72 hours following the transfection with a scramble siRNA (siGL3) and a siRNA targeting Ago1 (siAgo1).