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Medicina is published by MDPI from Volume 54 Issue 1 (2018). Articles in this Issue were published by another publisher in Open Access under a CC-BY (or CC-BY-NC-ND) licence. Articles are hosted by MDPI on as a courtesy and upon agreement with Lithuanian Medical Association, Lithuanian University of Health Sciences, and Vilnius University.
Open AccessArticle

High-Resolution Melting-Based Quantitative Analysis of RASSF1 Methylation in Breast Cancer

Division of Human Genome Research Centre, Faculty of Natural Sciences, Vilnius University
National Centre of Pathology, Affiliate of Vilnius University Hospital Santariškių Klinikos
Institute of Oncology, Vilnius University, Lithuania
Author to whom correspondence should be addressed.
Medicina 2013, 49(2), 14;
Received: 15 January 2013 / Accepted: 28 February 2013 / Published: 5 March 2013
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).
. 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.
. HRM is as a simple, cost-effective method for the reliable high-throughput quantification of DNA methylation in clinical material.
Keywords: DNA methylation; breast cancer; HRM; QMSP; RASSF1 gene DNA methylation; breast cancer; HRM; QMSP; RASSF1 gene
MDPI and ACS Style

Stuopelytė, K.; Daniūnaitė, K.; Laurinavičienė, A.; Ostapenko, V.; Jarmalaitė, S. High-Resolution Melting-Based Quantitative Analysis of RASSF1 Methylation in Breast Cancer. Medicina 2013, 49, 14.

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