Related Experiment Video
Updated: May 8, 2026
Methyl-binding DNA capture Sequencing for Patient Tissues
Published on: October 31, 2016
Quantitative evaluation of DNMT3B promoter methylation in breast cancer patients using differential high resolution
M Naghitorabi1, J Mohammadi Asl, H Mir Mohammad Sadeghi
1Department of Pharmaceutical Biotechnology and Isfahan Pharmaceutical Sciences Research Center School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.
Abstract:
DNA methylation plays an important role in carcinogenesis through epigenetic silencing of tumor suppressor genes. Aberrant methylation usually results from changes in the activity of DNA methyltransferases (DNMTs). Some studies show that the overexpression of the DNMTs may lead to aberrant methylation of tumor suppressor genes. Also the overexpression of DNMTs may be related to methylation status of their genes. Due to limited number of studies on DNMT3B promoter methylation, this study was performed to quantitatively measure the methylation level of DNMT3B gene in archival formalin fixed paraffin embedded (FFPE) tissues from breast cancer patients. Using differential high resolution melting analysis (D-HRMA) technology, the methylation level of DNMT3B gene promoter was quantified in 98 breast cancer FFPE tissues and also 10 fresh frozen normal tissue samples. Statistical analyses used for analyzing the correlation between the methylation and clinical variables. All the normal samples were found to be methylated at the DNMT3B promoter (the average methylation level 3.34%). Patients were identified as hypo-methylated (mean methylation level 0.8%), methylated (mean methylation level 2.48%) and hyper-methylated (mean methylation level 10.5%). Statistical analysis showed a significant correlation between the methylation status and the sample type, cancer type and tumor size. Also the methylation level was significantly associated with histologic grade. It is concluded that quantification of DNMT3B promoter methylation might be used as a reliable and sensitive diagnostic and prognostic tool in breast cancer. Also D-HRMA is demonstrated as a rapid and cost effective method for quantitative evaluation of promoter methylation.
Insights
DNA methylation is crucial in breast cancer. This study quantified DNMT3B promoter methylation in tumors, finding it correlates with cancer type, size, and grade, suggesting its diagnostic potential.
Area of Science:
- Epigenetics
- Cancer Biology
- Molecular Diagnostics
Background:
- DNA methylation is a key epigenetic mechanism in cancer, often involving DNA methyltransferases (DNMTs).
- Aberrant methylation, particularly of tumor suppressor genes, is linked to cancer development.
- Limited research exists on the methylation status of the DNMT3B gene promoter.
Purpose of the Study:
- To quantitatively measure the methylation level of the DNMT3B gene promoter in breast cancer tissues.
- To investigate the correlation between DNMT3B promoter methylation and clinical variables in breast cancer patients.
- To evaluate the potential of DNMT3B promoter methylation as a diagnostic and prognostic tool.
Main Methods:
- Quantitative analysis of DNMT3B promoter methylation using differential high resolution melting analysis (D-HRMA).
- Study included 98 breast cancer formalin-fixed paraffin-embedded (FFPE) tissues and 10 fresh frozen normal tissue samples.
- Statistical analyses were performed to correlate methylation status with clinical and histologic variables.
Main Results:
- All normal samples exhibited methylation at the DNMT3B promoter (average 3.34%).
- Breast cancer patients were categorized into hypo- (0.8%), methylated (2.48%), and hyper-methylated (10.5%) groups.
- Significant correlations were observed between DNMT3B methylation status and sample type, cancer type, tumor size, and histologic grade.
Conclusions:
- Quantification of DNMT3B promoter methylation may serve as a reliable diagnostic and prognostic biomarker in breast cancer.
- D-HRMA is presented as an efficient, rapid, and cost-effective method for assessing promoter methylation.
- Further research could validate DNMT3B methylation as a clinical tool for breast cancer management.
