title: Methylation pattern analysis in prostate cancer tissue: identification of biomarkers using an MS-MLPA approach.
abstract: Epigenetic silencing mediated by CpG island methylation is a common feature of many cancers. Characterizing aberrant DNA methylation changes associated with prostate carcinogenesis could potentially identify a tumour-specific methylation pattern, facilitating the early diagnosis of prostate cancer. The objective of the study was to assess the methylation status of 40 tumour suppressor genes in prostate cancer and healthy prostatic tissues.
  
  
  
  We used methylation specific-multiplex ligation probe amplification (MS-MLPA) assay in two independent case series (training and validation set). The training set comprised samples of prostate cancer tissue (n = 40), healthy prostatic tissue adjacent to the tumor (n = 26), and healthy non prostatic tissue (n = 23), for a total of 89 DNA samples; the validation set was composed of 40 prostate cancer tissue samples and their adjacent healthy prostatic tissue, for a total of 80 DNA samples. Methylation specific-polymerase chain reaction (MSP) was used to confirm the results obtained in the validation set.
  
  
  
  We identified five highly methylated genes in prostate cancer: GSTP1, RARB, RASSF1, SCGB3A1, CCND2 (P < 0.0001), with an area under the ROC curve varying between 0.89 (95 % CI 0.82-0.97) and 0.95 (95 % CI 0.90-1.00). Diagnostic accuracy ranged from 80 % (95 % CI 70-88) to 90 % (95 % CI 81-96). Moreover, a concordance rate ranging from 83 % (95 % CI 72-90) to 89 % (95 % CI 80-95) was observed between MS-MLPA and MSP.
  
  
  
  Our preliminary results highlighted that hypermethylation of GSTP1, RARB, RASSF1, SCGB3A1 and CCND2 was highly tumour-specific in prostate cancer tissue.
authors: Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Via P. Maroncelli 40, 47014, Meldola, Italy.
authors: Giorgia
authors: G
authors: Gurioli
authors: Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Via P. Maroncelli 40, 47014, Meldola, Italy.
authors: Samanta
authors: S
authors: Salvi
authors: Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Via P. Maroncelli 40, 47014, Meldola, Italy.
authors: Filippo
authors: F
authors: Martignano
authors: Unit of Biostatistics and Clinical Trials, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Via P. Maroncelli 40, 47014, Meldola, Italy.
authors: Flavia
authors: F
authors: Foca
authors: Department of Urology, Morgagni Pierantoni Hospital, Forlì, Italy.
authors: Roberta
authors: R
authors: Gunelli
authors: Pathology Unit, Morgagni Pierantoni Hospital, Forlì, Italy.
authors: Matteo
authors: M
authors: Costantini
authors: Department of Urology, Bufalini Hospital, Cesena, Italy.
authors: Giacomo
authors: G
authors: Cicchetti
authors: Department of Medical Oncology, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Via P. Maroncelli 40, 47014, Meldola, Italy.
authors: Ugo
authors: U
authors: De Giorgi
authors: Dipartimento di Scienze Biomediche, Sperimentali e Cliniche, University of Florence, Florence, Italy.
authors: Persio Dello
authors: PD
authors: Sbarba
authors: Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Via P. Maroncelli 40, 47014, Meldola, Italy.
authors: Daniele
authors: D
authors: Calistri
authors: Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Via P. Maroncelli 40, 47014, Meldola, Italy. valentina.casadio@irst.emr.it.
authors: Valentina
authors: V
authors: Casadio
mesh_terms: D000368:Aged; D014408:Biomarkers, Tumor / Q000235:genetics*; D016022:Case-Control Studies; D016000:Cluster Analysis; D019175:DNA Methylation / Q000235:genetics*; D016147:Genes, Tumor Suppressor; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011471:Prostatic Neoplasms / Q000175:diagnosis / Q000235:genetics*; D012372:ROC Curve; D015203:Reproducibility of Results
chemical_list: D014408:Biomarkers, Tumor
keywords: Early diagnosis; MS-MLPA; Methylation pattern; Prostate cancer