Rapid and reliable detection of LINE-1 hypomethylation using high-resolution melting analysis

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• 作者：Stefanie Stanzer ; Marija Balic ; Jasmin Strutz ; Ellen Heitzer ; Florian Obermair ; Cornelia Hauser-Kronberger ; Hellmut Samonigg ; Nadia D ; achi
• 关键词：Hypomethylation ; LINE-1 ; Repetitive elements ; High-resolution melting ; MethyLight ; Pyrosequencing ; Archival prostate tumor samples
• 刊名：Clinical Biochemistry
• 出版年：2010
• 出版时间：December 2010
• 年：2010
• 卷：43
• 期：18
• 页码：1443-1448
• 全文大小：567 K

文摘

Objectives

The aim of the present study was to evaluate the precision and reproducibility of the LINE-1 high-resolution melting (HRM) assay to detect LINE-1 hypomethylation.

Design and methods

We first evaluated a methylated DNA dilution matrix and a panel of human cancer cell lines. We then applied this LINE-1 HRM assay to a set of 37 archival prostate cancer tissue samples.

Results

Our LINE-1 HRM assay revealed small and reproducible run-to-run and bisulfite-to-bisulfite variations. As expected, we found a large variation in methylation levels between different cancer cell lines. All results were confirmed with MethyLight and pyrosequencing as indicated by the high correlation coefficient. Finally, we successfully applied the LINE-1 HRM assay to archival prostate cancer tissues.

Conclusions

The present LINE-1 HRM assay represents a novel, accurate, and cost-effective method to measure global hypomethylation, which makes it suitable for high- and low-throughput laboratories.