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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 mdpi.com 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

1
Division of Human Genome Research Centre, Faculty of Natural Sciences, Vilnius University
2
National Centre of Pathology, Affiliate of Vilnius University Hospital Santariškių Klinikos
3
Institute of Oncology, Vilnius University, Lithuania
*
Author to whom correspondence should be addressed.
Medicina 2013, 49(2), 14; https://doi.org/10.3390/medicina49020014
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).
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.
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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