title: miR-204 is dysregulated in metastatic prostate cancer in vitro.
abstract: During cancer progression, the genome instability incurred rearrangement could possibly turn some of the tumor suppressor micro-RNAs into pro-oncogenic ones. We aimed to investigate miR-204 in the context of prostate cancer progression using a cell line model of different levels of genome instability (LNCaP, PC3, VCaP and NCI H660), as demonstrated by the availability of ERG fusion. We studied the effect of miR-204 modulation on master transcription factors important for lineage development, cell differentiation and prostate cancer bone marrow metastasis. We followed c-MYB, ETS1 and RUNX2 transcript and protein expression and the miR-204 affected global proteome. We further investigated if these transcription factors exert an effect on miR-204 expression (qPCR, luciferase reporter assay) by silencing them using esiRNA. We found dualistic miR-204 effects, either acting as a tumor suppressor on c-MYB, or as an oncomiR on ETS1. RUNX2 and ETS1 regulation by miR-204 was ERG fusion dependent, demonstrating regulatory circuitry disruption in advanced metastatic models. miR-204 also differentially affected mRNA splicing and protein stability. miR-204 levels were found dependent on cancer hypermethylation and supported by positive feedback induced by all three transcription factors. In this regulatory circuitry among miR-204, c-MYB, RUNX2 and ETS1, the c-MYB was found to induce all three other members, but its expression was differentially affected by the methylation status in lymph node vs. bone metastasis. We demonstrate that not only tumor suppressor micro-RNA loss, but also significant genome rearrangement-driven regulatory loop perturbations play a role in the advanced cancer progression, conferring better pro-survival and metastatic potential.
authors: Institute of Biology and Immunology of Reproduction at Bulgarian Academy of Sciences, Sofia, Bulgaria.
authors: Krassimira
authors: K
authors: Todorova
authors: School of Biological Sciences, Essex University, Colchester, UK.
authors: Metodi V
authors: MV
authors: Metodiev
authors: School of Biological Sciences, Essex University, Colchester, UK.
authors: Gergana
authors: G
authors: Metodieva
authors: Institute of Biology and Immunology of Reproduction at Bulgarian Academy of Sciences, Sofia, Bulgaria.
authors: Diana
authors: D
authors: Zasheva
authors: Cellular and Gene Therapy Ward, National Specialized Hematology Hospital, Sofia, Bulgaria.
authors: Milcho
authors: M
authors: Mincheff
authors: Institute of Biology and Immunology of Reproduction at Bulgarian Academy of Sciences, Sofia, Bulgaria.
authors: Soren
authors: S
authors: Hayrabedyan
mesh_terms: D017398:Alternative Splicing; D045744:Cell Line, Tumor; D050659:Core Binding Factor Alpha 1 Subunit / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D009362:Neoplasm Metastasis; D015514:Oncogene Proteins, Fusion / Q000235:genetics; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D055550:Protein Stability; D020543:Proteome / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D050784:Proto-Oncogene Protein c-ets-1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020598:Proto-Oncogene Proteins c-myb / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics; D000071230:Transcriptional Regulator ERG
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; C072312:ERG protein, human; C494041:ETS1 protein, human; C549404:MIRN204 microRNA, human; D035683:MicroRNAs; D015514:Oncogene Proteins, Fusion; D020543:Proteome; D050784:Proto-Oncogene Protein c-ets-1; D020598:Proto-Oncogene Proteins c-myb; C493709:RUNX2 protein, human; D015534:Trans-Activators; D000071230:Transcriptional Regulator ERG
keywords: (MeSH); DNA Methylation; LTQ Orbitrap Velos Proteomics; Micro RNA; Oncogene Fusion; Transcription Factor