Hypermethylation of the GABRE∼miR-452∼miR-224 Promoter in Prostate Cancer Predicts Biochemical Recurrence after Radical Prostatectomy

Purpose: Available tools for prostate cancer diagnosis and prognosis are suboptimal and novel biomarkers are urgently needed. Here, we investigated the regulation and biomarker potential of the GABRE∼miR-452∼miR-224 genomic locus. Experimental Design: GABRE/miR-452/miR-224 transcriptional expression was quantified in 80 nonmalignant and 281 prostate cancer tissue samples. GABRE∼miR-452∼miR-224 promoter methylation was determined by methylation-specific qPCR (MethyLight) in 35 nonmalignant, 293 prostate cancer [radical prostatectomy (RP) cohort 1] and 198 prostate cancer tissue samples (RP cohort 2). Diagnostic/prognostic biomarker potential of GABRE∼miR-452∼miR-224 methylation was evaluated by ROC, Kaplan–Meier, uni- and multivariate Cox regression analyses. Functional roles of miR-224 and miR-452 were investigated in PC3 and DU145 cells by viability, migration, and invasion assays and gene-set enrichment analysis (GSEA) of posttransfection transcriptional profiling data. Results: GABRE∼miR-452∼miR-224 was significantly downregulated in prostate cancer compared with nonmalignant prostate tissue and had highly cancer-specific aberrant promoter hypermethylation (AUC = 0.98). Functional studies and GSEA suggested that miR-224 and miR-452 inhibit proliferation, migration, and invasion of PC3 and DU145 cells by direct/indirect regulation of pathways related to the cell cycle and cellular adhesion and motility. Finally, in uni- and multivariate analyses, high GABRE∼miR-452∼miR-224 promoter methylation was significantly associated with biochemical recurrence in RP cohort 1, which was successfully validated in RP cohort 2. Conclusion: The GABRE∼miR-452∼miR-224 locus is downregulated and hypermethylated in prostate cancer and is a new promising epigenetic candidate biomarker for prostate cancer diagnosis and prognosis. Tumor-suppressive functions of the intronic miR-224 and miR-452 were demonstrated in two prostate cancer cell lines, suggesting that epigenetic silencing of GABRE∼miR-452∼miR-224 may be selected for in prostate cancer. Clin Cancer Res; 20(8); 2169–81. ©2014 AACR.

[1]  N. Seki,et al.  The tumor-suppressive microRNA-143/145 cluster inhibits cell migration and invasion by targeting GOLM1 in prostate cancer , 2013, Journal of Human Genetics.

[2]  K. D. Sørensen,et al.  DNA methylation signatures for prediction of biochemical recurrence after radical prostatectomy of clinically localized prostate cancer. , 2013, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[3]  K. D. Sørensen,et al.  Prognostic significance of aberrantly silenced ANPEP expression in prostate cancer , 2013, British Journal of Cancer.

[4]  Dario Strbenac,et al.  Regional activation of the cancer genome by long-range epigenetic remodeling. , 2013, Cancer cell.

[5]  A. Scorilas,et al.  Downregulation and prognostic performance of microRNA 224 expression in prostate cancer. , 2013, Clinical chemistry.

[6]  Haibo Zhang,et al.  Expression of gamma-aminobutyric acid receptors on neoplastic growth and prediction of prognosis in non-small cell lung cancer , 2013, Journal of Translational Medicine.

[7]  A. D. de Carvalho,et al.  Positivity for HLA DR1 is associated with basal cell carcinoma in renal transplant patients in southern Brazil , 2012, International journal of dermatology.

[8]  Stefan Hüttelmaier,et al.  Insulin-like growth factor 2 mRNA-binding proteins (IGF2BPs): post-transcriptional drivers of cancer progression? , 2012, Cellular and Molecular Life Sciences.

[9]  M. Kuczyk,et al.  Evaluating the significance of cyclic adenosine monophosphate-mediated signaling in human prostate: a functional and biochemical study. , 2012, Urology.

[10]  Gang Chen,et al.  Diagnostic significance of overexpression of Golgi membrane protein 1 in prostate cancer. , 2012, Urology.

[11]  Huiyong Shen,et al.  MicroRNA-224 targets RKIP to control cell invasion and expression of metastasis genes in human breast cancer cells. , 2012, Biochemical and biophysical research communications.

[12]  B. Trock,et al.  Evaluation of GSTP1 and APC methylation as indicators for repeat biopsy in a high‐risk cohort of men with negative initial prostate biopsies , 2012, BJU international.

[13]  K. Kelnar,et al.  Distinct microRNA expression profiles in prostate cancer stem/progenitor cells and tumor-suppressive functions of let-7. , 2012, Cancer research.

[14]  John T. Wei,et al.  Beyond PSA: The Next Generation of Prostate Cancer Biomarkers , 2012, Science Translational Medicine.

[15]  T. Visakorpi,et al.  Diagnostic and prognostic signatures from the small non-coding RNA transcriptome in prostate cancer , 2012, Oncogene.

[16]  Sung-Wuk Jang,et al.  Regulation of ADP-ribosylation factor 4 expression by small leucine zipper protein and involvement in breast cancer cell migration. , 2012, Cancer letters.

[17]  T. Rohan,et al.  Hsa‐miR‐375 is differentially expressed during breast lobular neoplasia and promotes loss of mammary acinar polarity , 2012, The Journal of pathology.

[18]  S. Tavazoie,et al.  A microRNA regulon that mediates endothelial recruitment and metastasis by cancer cells , 2011, Nature.

[19]  W. Schulz,et al.  DNA methylation changes in prostate cancer. , 2012, Methods in molecular biology.

[20]  Agnieszka Piekielko-Witkowska,et al.  MiR-224 Targets the 3′UTR of Type 1 5′-Iodothyronine Deiodinase Possibly Contributing to Tissue Hypothyroidism in Renal Cancer , 2011, PloS one.

[21]  N. Seki,et al.  Restoration of miR-145 expression suppresses cell proliferation, migration and invasion in prostate cancer by targeting FSCN1. , 2011, International journal of oncology.

[22]  Wei Wu.  MicroRNA and Cancer , 2011, Methods in Molecular Biology.

[23]  K. D. Sørensen,et al.  Promoter hypomethylation and upregulation of trefoil factors in prostate cancer , 2010, International journal of cancer.

[24]  Aliasgar Moiyadi,et al.  Distinctive microRNA signature of medulloblastomas associated with the WNT signaling pathway. , 2010, Journal of cancer research and therapeutics.

[25]  C. Sander,et al.  Integrative genomic profiling of human prostate cancer. , 2010, Cancer cell.

[26]  S. Freedland,et al.  Multicenter clinical validation of PITX2 methylation as a prostate specific antigen recurrence predictor in patients with post-radical prostatectomy prostate cancer. , 2010, The Journal of urology.

[27]  D. Geerts,et al.  The polyamine metabolism genes ornithine decarboxylase and antizyme 2 predict aggressive behavior in neuroblastomas with and without MYCN amplification , 2010, International journal of cancer.

[28]  G. Yousef,et al.  Three dysregulated miRNAs control kallikrein 10 expression and cell proliferation in ovarian cancer , 2010, British Journal of Cancer.

[29]  Zhimin Zhang,et al.  MicroRNA‐224 is upregulated in HepG2 cells and involved in cellular migration and invasion , 2010, Journal of gastroenterology and hepatology.

[30]  Jianfeng Xu,et al.  Open Access RESEARCH , 2010 .

[31]  Huiqing Liu,et al.  Identifying mRNA targets of microRNA dysregulated in cancer: with application to clear cell Renal Cell Carcinoma , 2010, BMC Systems Biology.

[32]  M. Lucia,et al.  Prostate Cancer Detected by Methylated Gene Markers in Histopathologically Cancer-Negative Tissues from Men with Subsequent Positive Biopsies , 2009, Cancer Epidemiology, Biomarkers & Prevention.

[33]  R. Kooy,et al.  The complexity of the GABAA receptor shapes unique pharmacological profiles. , 2009, Drug discovery today.

[34]  Å. Borg,et al.  MiRNA expression in urothelial carcinomas: Important roles of miR‐10a, miR‐222, miR‐125b, miR‐7 and miR‐452 for tumor stage and metastasis, and frequent homozygous losses of miR‐31 , 2009, International journal of cancer.

[35]  Chang Woo Han,et al.  Identification of ADP-ribosylation factor 4 as a suppressor of N-(4-hydroxyphenyl)retinamide-induced cell death. , 2009, Cancer letters.

[36]  G. Kristiansen,et al.  DNA methylation of the PITX2 gene promoter region is a strong independent prognostic marker of biochemical recurrence in patients with prostate cancer after radical prostatectomy. , 2009, The Journal of urology.

[37]  A. Bittner,et al.  Characterization of global microRNA expression reveals oncogenic potential of miR-145 in metastatic colorectal cancer , 2009, BMC Cancer.

[38]  Gerhard K. H. Przemeck,et al.  The CALM and CALM/AF10 interactor CATS is a marker for proliferation , 2008, Molecular oncology.

[39]  S. Dhanasekaran,et al.  Golgi protein GOLM1 is a tissue and urine biomarker of prostate cancer. , 2008, Neoplasia.

[40]  C. Pilarsky,et al.  GOLPH2 protein expression as a novel tissue biomarker for prostate cancer: implications for tissue-based diagnostics , 2008, British Journal of Cancer.

[41]  George A. Calin,et al.  Expression of microRNAs and protein‐coding genes associated with perineural invasion in prostate cancer , 2008, The Prostate.

[42]  Anne-Mette K. Hein,et al.  Alternative Splicing in Colon, Bladder, and Prostate Cancer Identified by Exon Array Analysis*S , 2008, Molecular & Cellular Proteomics.

[43]  Jianjun Yu,et al.  A first-generation multiplex biomarker analysis of urine for the early detection of prostate cancer. , 2008, Cancer research.

[44]  N. Hoosein,et al.  Expression of gamma-aminobutyric acid receptor (subtype A) in prostate cancer , 2008, Acta oncologica.

[45]  J. Cheng,et al.  Identification of miRNAs associated with tumorigenesis of retinoblastoma by miRNA microarray analysis , 2008, Child's Nervous System.

[46]  R. Stephens,et al.  Unique microRNA molecular profiles in lung cancer diagnosis and prognosis. , 2006, Cancer cell.

[47]  A. Houtsmuller,et al.  A bioinformatics-based functional analysis shows that the specifically androgen-regulated gene SARG contains an active direct repeat androgen response element in the first intron. , 2004, Journal of molecular endocrinology.

[48]  T. L. Drell,et al.  Effects of Neurotransmitters on the Chemokinesis and Chemotaxis of MDA-MB-468 Human Breast Carcinoma Cells , 2003, Breast Cancer Research and Treatment.

[49]  K. Takahara,et al.  γ-Aminobutyric Acid as a Promoting Factor of Cancer Metastasis; Induction of Matrix Metalloproteinase Production Is Potentially Its Underlying Mechanism , 2003 .

[50]  M. Bianchi,et al.  Spontaneous and gamma-aminobutyric acid (GABA)-activated GABA(A) receptor channels formed by epsilon subunit-containing isoforms. , 1999, Molecular pharmacology.

[51]  S. Klauck,et al.  A gene in human chromosome band Xq28 (GABRE) defines a putative new subunit class of the GABAA neurotransmitter receptor. , 1997, Genomics.