High Quality Assessment of DNA Methylation in Archival Tissues from Colorectal Cancer Patients Using Quantitative High-Resolution Melting Analysis

被引：55

作者：

Balic, Marija ^{[1
]}

Pichler, Martin ^{[1
]}

Strutz, Jasmin ^{[1
]}

Heitzer, Ellen ^{[1
]}

Ausch, Christoph ^{[2
]}

Samonigg, Hellmut ^{[1
]}

Cote, Richard J. ^{[3
]}

Dandachi, Nadia ^{[1
]}

机构：

[1] Med Univ Graz, Dept Internal Med, Div Oncol, A-8036 Graz, Austria

[2] Danube Hosp, Ludwig Boltzmann Res Inst Surg Oncol, Dept Surg, Vienna, Austria

[3] Univ So Calif, Keck Sch Med, Dept Pathol, Los Angeles, CA 90033 USA

来源：

JOURNAL OF MOLECULAR DIAGNOSTICS | 2009年 / 11卷 / 02期

关键词：

LYMPH-NODE METASTASES; PROSTATE-CANCER; BREAST-CANCER; PROMOTER HYPERMETHYLATION; GENE; PCR; INVASION; DENSITY; MARKERS;

D O I：

10.2353/jmoldx.2009.080109

中图分类号：

R36 [病理学];

学科分类号：

100104 ;

摘要：

High-resolution melting (HRM) analysis is a novel tool for analysis of promoter methylation. The aim of the present study was to establish and validate HRM analysis for detection of promoter methylation on archival formatin-fixed paraffin-embedded tissues from colorectal cancer patients. We first evaluated HRM assays for O(6)-methylguanine-DNA methyltransferase (MGMT) and adenomatous polyposis coli (APC) promoter methylation on a methylated DNA dilution matrix and DNA extracted from eight fresh or formalin-fixed paraffin-embedded human cancer cell fines. Then we used these assays for the analysis of MGMT and APC promoter methylation in a subset of archival formatin-fixed. paraffin-embedded colorectal tumor specimens. All samples with promoter methylation of MGMT or APC and randomly selected samples without promoter methylation were analyzed twice. All results generated by HRM were validated with MGMT and APC MethyLight assays. APC andMGMT promoter methylation data were consistent and reproducible throughout the dilutions and all three replicates in the methylated DNA dilution matrix and between two experiments in clinical samples. There was high concordance between HRM and MethyLight results. HRM for APC promoter methylation revealed consistent results between fresh and formalin-fixed paraffin-embedded human cancer cell line DNA. The methylation status in archival tumor specimens from patients with colorectal cancer can therefore be determined with high quality by HRM. The ability to analyze archival tissues greatly facilitates further research and its clinical implementation. (J Mol Diagn 2008,11:102-108 DOI: 10.2353/jmoldx.2009.080109)

引用

页码：102 / 108

页数：7

共 32 条

[1] APC promoter hypermethylation contributes to the loss of APC expression in colorectal cancers with allelic loss on 5q [J].