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Abstract
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. 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.
Highlights
Introduction the detection ofBCa at early stages and reduce the Breast cancer (BCa) is the most prevalent and risk of disease progression and mortality rates.leading cause of death from malignant diseas- Carcinogenesis overwhelms all cellular processes among women in Lithuania and worldwide
A total of 76 breast carcinomas and 10 noncancerous breast tissues were studied by means of high-resolution melting (HRM) and compared with the results obtained by means of quantitative methylation-specific polymerase chain reaction (QMSP) and methylation-specific polymerase chain reaction (MSP)
The mean methylation intensity of RASSF1 was 42.5% and 48.6% according to HRM and QMSP, respectively
Summary
Introduction the detection ofBCa at early stages and reduce the Breast cancer (BCa) is the most prevalent and risk of disease progression and mortality rates.leading cause of death from malignant diseas- Carcinogenesis overwhelms all cellular processes among women in Lithuania and worldwide. Invention of the quantitaprogesterone receptor (PR), and epidermal growth tive means of DNA methylation measurement enfactor receptor 2 (HER2), and the expression of abled the identification of minor variations in the cell proliferation marker Ki67 are used in diagnos- levels of methylation among tumors with different tics and prognostics of BCa [2]. According to these aggressiveness and provided a new tool for therapy biomarkers, BCa is divided into biological subtypes; individualization. The genome-wide profiling of gene material with the different density of cancer cells
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