ArticleClinical epigenetics2016
Detection and monitoring of hypermethylated RASSF1A in serum from patients with metastatic breast cancer.
Article in Clinical epigenetics, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers, 1 of them a synthesis that pooled it.
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Who cites it
10 citing papers in PubMed, 1 synthesis or guideline pooled it, 17 citations in OpenAlex.
- Liquid biopsy prediction of axillary lymph node metastasis, cancer recurrence, and patient survival in breast cancer: A meta-analysis.Medicine · 2018Pooled it
- Tumor and serum DNA methylation in women receiving preoperative chemotherapy with or without vorinostat in TBCRC008.Breast cancer research and treatment · 2018Trial
- Νovel methylation biomarkers in liquid biopsy and classifying biosignatures for the clinical management of breast cancer.Breast cancer research : BCR · 2026Article
- Liquid biopsy in breast cancer: Redefining precision medicine.The journal of liquid biopsy · 2025Review
- Prediction of blood-based biomarkers and subsequent design of bisulfite PCR-LDR-qPCR assay for breast cancer detection.BMC cancer · 2020Article
- Highly sensitive detection of sentinel lymph node metastasis of breast cancer by digital PCR for RASSF1A methylation.Oncology reports · 2019Article
- Next-generation sequencing in liquid biopsy: cancer screening and early detection.Human genomics · 2019Review
- Personalized medicine in breast cancer: pharmacogenomics approaches.Pharmacogenomics and personalized medicine · 2019Review
- Electrochemical affinity biosensors for fast detection of gene-specific methylations with no need for bisulfite and amplification treatments.Scientific reports · 2018Article
- Value of the level of methylation of RASSF1A and WIF-1 in tissue and serum in neoadjuvant chemotherapeutic assessment for advanced breast cancer.Oncology letters · 2017Article
Corrections and comments
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Authors and funding
3 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundCirculating hypermethylated RASSF1A could be a novel and potential useful marker for monitoring patients with metastatic breast cancer. Technical obstacles include fragmentation of the circulating DNA, fluctuations in the concentration, low concentrations of circulating tumor DNA, and different locations of methylation in the RASSF1A gene among patients. One common method for detection of hypermethylated genes is sodium bisulfite conversion of non-methylated cytosine to uracil, followed by detection with PCR. However, the method relies on full conversion of all non-methylated cytosines, cause strand breaks, and loss of DNA. Alternatively, methylation-sensitive restriction enzymes have been used to digest genomic DNA, as well as sodium bisulfite-treated DNA. By flanking different regions of the RASSF1A with different PCR primer pairs, we analyzed for methylated genomic regions resistant to cleavage by the methylation-sensitive restriction enzymes HpaII and BstUI. The goal was to find region(s) in RASSF1A with high sensitivity and specificity that could be used for monitoring.
resultsThe serum was spiked with non-human control DNA. By tracing the spiking control, the isolation procedure of the rare circulating tumor DNA was initially optimized. By analysis of production of PCR amplicons from HpaII- or BstUI-treated DNA isolated from 24 patients with metastatic breast cancer, we located four regions resulting in sensitivities from 63 to 83 %. When examining samples from 24 control subjects, these four regions gave a specificity of 100 %. Among these four regions, the primer pair with the highest PCR efficacy was selected to monitor the RASSF1A concentration in 31 collected serum samples. The spiked DNA was then used to calculate the tumor RASSF1A concentrations independent of fluctuations in circulating non-tumor DNA. As a proof of principle, there was concordance in the kinetics of the RASSF1A and the serological cancer biomarkers CA 15-3, CEA, and TPA.
conclusionsMethylation-sensitive restriction enzymes may be a useful methodological approach for monitoring circulating hypermethylated RASSF1A among patients with metastatic breast cancer.
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