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Detection and monitoring of hypermethylated RASSF1A in serum from patients with metastatic breast cancer

Overview of attention for article published in Clinical Epigenetics, April 2016
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Title
Detection and monitoring of hypermethylated RASSF1A in serum from patients with metastatic breast cancer
Published in
Clinical Epigenetics, April 2016
DOI 10.1186/s13148-016-0199-0
Pubmed ID
Authors

Søren Kristiansen, Dorte Nielsen, György Sölétormos

Abstract

Circulating 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. The 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. Methylation-sensitive restriction enzymes may be a useful methodological approach for monitoring circulating hypermethylated RASSF1A among patients with metastatic breast cancer.

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Mendeley readers

The data shown below were compiled from readership statistics for 33 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Libya 1 3%
United States 1 3%
Unknown 31 94%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 6 18%
Student > Bachelor 4 12%
Researcher 4 12%
Student > Master 4 12%
Professor > Associate Professor 3 9%
Other 7 21%
Unknown 5 15%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 8 24%
Medicine and Dentistry 7 21%
Agricultural and Biological Sciences 5 15%
Chemistry 2 6%
Nursing and Health Professions 1 3%
Other 4 12%
Unknown 6 18%

Attention Score in Context

This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 12 April 2016.
All research outputs
#9,318,463
of 15,876,641 outputs
Outputs from Clinical Epigenetics
#504
of 839 outputs
Outputs of similar age
#128,896
of 266,790 outputs
Outputs of similar age from Clinical Epigenetics
#10
of 13 outputs
Altmetric has tracked 15,876,641 research outputs across all sources so far. This one is in the 39th percentile – i.e., 39% of other outputs scored the same or lower than it.
So far Altmetric has tracked 839 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 5.7. This one is in the 36th percentile – i.e., 36% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 266,790 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 48th percentile – i.e., 48% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 13 others from the same source and published within six weeks on either side of this one. This one is in the 7th percentile – i.e., 7% of its contemporaries scored the same or lower than it.