Quantitative assessment of AKAP12 promoter methylation in human prostate cancer using methylation-sensitive high-resolution melting: correlation with Gleason score.

Abstract

OBJECTIVES:

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: Weiwei Liu, Jian Gong, Jinghui Hu, Tingting Hu, Yaofei Sun, Junhua Du, Chuanyu Sun, Ming Guan, Haowen Jiang, Yuan Lu
• Journal: Urology (Urology) Vol. 77 Issue 4 Pg. 1006.e1-7 (Apr 2011) ISSN: 1527-9995 [Electronic] United States
• PMID: 21310466 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2011 Elsevier Inc. All rights reserved.

Chemical References

• A Kinase Anchor Proteins
• AKAP12 protein, human
• Cell Cycle Proteins
• Mitogens
• Decitabine
• DNA Modification Methylases
• Azacitidine

Topics

• A Kinase Anchor Proteins (chemistry, genetics)
• Azacitidine (analogs & derivatives, pharmacology)
• Cell Cycle Proteins (chemistry, genetics)
• Cell Line, Tumor (drug effects)
• DNA Methylation
• DNA Modification Methylases (antagonists & inhibitors)
• Decitabine
• Humans
• Male
• Mitogens (chemistry, genetics)
• Promoter Regions, Genetic (genetics)
• Prostatic Hyperplasia (genetics)
• Prostatic Neoplasms (metabolism, pathology)
• Reverse Transcriptase Polymerase Chain Reaction