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Medicina (Kaunas). 2013;49(2):78-83.

High-resolution melting-based quantitative analysis of RASSF1 methylation in breast cancer.

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1
Division of Human Genome Research Centre, Faculty of Natural Sciences, Vilnius University, M. K. Čiurlionio 21, 03101 Vilnius, Lithuania. sonata.jarmalaite@gf.vu.lt

Abstract

BACKGROUND AND OBJECTIVE:

Breast cancer is the leading cause of death from cancer among women worldwide. The aberrant promoter methylation of tumor suppressor genes is a typical epigenetic alteration for breast cancer and can be detected in early carcinogenesis. High-throughput and cost-effective methods are needed for the early and sensitive detection of epigenetic changes in clinical material. The main purpose of our study was to optimize a high-resolution melting (HRM) assay for the reliable and quantitative assessment of RASSF1 gene methylation, which is considered one of the earliest epigenetic alterations in breast cancer.

MATERIAL AND METHODS:

A total of 76 breast carcinomas and 10 noncancerous breast tissues were studied by means of HRM and compared with the results obtained by means of quantitative methylation-specific polymerase chain reaction (QMSP) and methylation-specific polymerase chain reaction (MSP).

RESULTS:

Both quantitative methods, HRM and QMSP, showed a similar specificity and sensitivity for the detection of RASSF1 methylation in breast cancer (about 80% and 70%, respectively). In breast cancer, the mean methylation intensity of RASSF1 was 42.5% and 48.6% according to HRM and QMSP, respectively. Both methods detected low levels of methylation (less than 5%) in noncancerous breast tissues. In comparison with quantitative methods, MSP showed a lower sensitivity (70%), but a higher specificity (80%) for the detection of RASSF1 methylation in breast cancer.

CONCLUSIONS:

HRM is as a simple, cost-effective method for the reliable high-throughput quantification of DNA methylation in clinical material.

PMID:
23888343
[Indexed for MEDLINE]
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