title: Quantitative assessment of AKAP12 promoter methylation in human prostate cancer using methylation-sensitive high-resolution melting: correlation with Gleason score.
abstract: OBJECTIVE
  To quantitatively investigate the A kinase anchoring protein 12 (AKAP12) gene promoter methylation and its association with clinicopathologic variables in human prostate cancer (PCa). The AKAP12 gene has shown reduced expression and marked hypermethylation in a variety of cancers.
  
  
  METHODS
  The percentage levels of DNA methylation were measured in 78 PCa, 22 benign prostatic hyperplasia, and 22 normal adjacent tissue samples using an AKAP12 methylation-sensitive high-resolution melting assay. AKAP12 gene expression was also examined in 4 human prostate carcinoma cell lines, PC-3, DU145, LNCaP, and 22RV1, using quantitative reverse transcriptase-polymerase chain reaction and methylation-sensitive high-resolution melting analysis and after DNA methyltransferase inhibition with 5-aza-2'-deoxycytidine.
  
  
  RESULTS
  Methylation (>1%) of the AKAP12 promoter region was present in 47 (60.2%) of the 78 PCa, 5 (22.7%) of the 22 benign prostatic hyperplasia, and 2 (9.1%) of the 22 adjacent normal tissue samples. AKAP12 methylation was significantly greater in the PCa than in the benign prostatic hyperplasia or adjacent tissue samples (P < .01). AKAP12 methylation was significantly greater in the PCa samples with higher Gleason scores (P = .03); however, no correlation was found with age, pT category, or serum prostate-specific antigen level. Reverse transcriptase-polymerase chain reaction demonstrated that PC-3 and DU-145 cells expressed AKAP12 RNA and LNCaP and 22RV1 did not. The AKAP12 locus was methylated in the LNCaP and 22RV1 cells. Treatment of LNCaP cells with 5-aza-2'-deoxycytidine markedly decreased the methylation levels and increased the expression of AKAP12.
  
  
  CONCLUSIONS
  The results of the present study have demonstrated that AKAP12 promoter methylation is a frequent event in human PCa. AKAP12 methylation represents a potential molecular biomarker for predicting the malignancy of PCa.
authors.affiliation: Department of Laboratory Medicine, Huashan Hospital, Fudan University Shanghai Medical College, Shanghai, China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Liu
authors.forename: Jian
authors.initials: J
authors.lastname: Gong
authors.forename: Jinghui
authors.initials: J
authors.lastname: Hu
authors.forename: Tingting
authors.initials: T
authors.lastname: Hu
authors.forename: Yaofei
authors.initials: Y
authors.lastname: Sun
authors.forename: Junhua
authors.initials: J
authors.lastname: Du
authors.forename: Chuanyu
authors.initials: C
authors.lastname: Sun
authors.forename: Ming
authors.initials: M
authors.lastname: Guan
authors.forename: Haowen
authors.initials: H
authors.lastname: Jiang
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lu
mesh_terms: D054758:A Kinase Anchor Proteins / Q000737:chemistry* / Q000235:genetics*; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000235:genetics*; D045744:Cell Line, Tumor / Q000187:drug effects; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors; D000077209:Decitabine; D006801:Humans; D008297:Male; D008934:Mitogens / Q000737:chemistry* / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011470:Prostatic Hyperplasia / Q000235:genetics; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D054758:A Kinase Anchor Proteins; C491162:AKAP12 protein, human; D018797:Cell Cycle Proteins; D008934:Mitogens; D000077209:Decitabine; D015254:DNA Modification Methylases; D001374:Azacitidine