Micro-RNA-204 Participates in TMPRSS2/ERG Regulation and Androgen Receptor Reprogramming in Prostate Cancer
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Cancer progression is driven by genome instability incurred rearrangements such as transmembrane protease, serine 2 (TMPRSS2)/v-ets erythroblastosis virus E26 oncogene (ERG) that could possibly turn some of the tumor suppressor micro-RNAs into pro-oncogenic ones. Previously, we found dualistic miR-204 effects, acting either as a tumor suppressor or as an oncomiR in ERG fusion-dependent manner. Here, we provided further evidence for an important role of miR-204 for TMPRSS2/ERG and androgen receptor (AR) signaling modulation and fine tuning that prevents TMPRSS2/ERG overexpression in prostate cancer. Based on proximity-based ligation assay, we designed a novel method for detection of TMPRSS2/ERG protein products. We found that miR-204 is TMPRSS2/ERG oncofusion negative regulator, and this was mediated by DNA methylation of TMPRSS2 promoter. Transcriptional factors runt-related transcription factor 2 (RUNX2) and ETS proto-oncogene 1 (ETS1) were positive regulators of TMPRSS2/ERG expression and promoter hypo-methylation. Clustering of patients’ sera for fusion protein, transcript expression, and wild-type ERG transcript isoforms, demonstrated not all patients harboring fusion transcripts had fusion protein products, and only few fusion positive ones exhibited increased wild-type ERG transcripts. miR-204 upregulated AR through direct promoter hypo-methylation, potentiated by the presence of ERG fusion and RUNX2 and ETS1. Proteomics studies provided evidence that miR-204 has dualistic role in AR cancer-related reprogramming, promoting prostate cancer-related androgen-responsive genes and AR target genes, as well as AR co-regulatory molecules. miR-204 methylation regulation was supported by changes in molecules responsible for chromatin remodeling, DNA methylation, and its regulation. In summary, miR-204 is a mild regulator of the AR function during the phase of preserved AR sensitivity as the latter one is required for ERG-fusion translocation.
KeywordsAndrogen Receptor LNCaP Cell Androgen Receptor Expression Proximity Ligation Assay Androgen Receptor Signaling
This study was funded by grants from The National Sciences Fund at Bulgarian Ministry of Education and Sciences (DMU 03/27 and DCOST 01/23) provided to Krassimira Todorova, Ph.D., D.Sc. and Soren Hayrabedyan, M.D., Ph.D., D.Sc.
The experimental work described herein was performed in laboratories of Institute of Biology and Immunology of Reproduction and Proteomics Core of Essex University, School of Biological Sciences. Method of detection of TMPRSS2/ERG protein products has patent pending procedure at Bulgarian National Patent Office (2014, K.T., S.H.).
K.T and S.H. designed the study, TMPRSS2/ERG protein detection method. K.T. did the cell-based experiments, FCS, gene silencing. and molecular biology studies. M.V.M. and G.M. did the LTQ Orbitrap proteomics and proteomics data analysis. S.H. did the bioinformatics analysis. M.M. and N.F. revised and refined the manuscript, providing constructive feedback.
Compliance with Ethical Standards
Conflict of Interest
The authors declare no conflict of interest.
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