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ID 52119
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Author
Muraoka, Takayuki
Toyooka, Shinichi
Aoe, Keisuke
Fujimoto, Nobukazu
Hashida, Shinsuke
Maki, Yuho
Tanaka, Norimitsu
Furukawa, Masashi
Yamamoto, Hiromasa ORCID Kaken ID researchmap
Kishimoto, Takumi
Otsuki, Takemi
Miyoshi, Shinichiro Kaken ID researchmap
Abstract
Objectives: Malignant pleural mesothelioma (MPM) is an aggressive tumor with a poor prognosis. microRNA-34b/c (miR-34b/c), which plays an important role in the pathogenesis of MPM, is frequently downregulated by DNA methylation in approximately 90% of MPM cases. In this study, we estimated the degree of miR-34b/c methylation in serum-circulating DNA using a digital methylation specific PCR assay (MSP). Materials and methods: A real-time MSP assay was performed using the SYBR Green method. The melting temperature (Tm) of each PCR product was examined using a melting curve analysis. For a digital MSP assay, 40 wells were analyzed per sample. A total of 110 serum samples from 48 MPM cases, 21 benign asbestos pleurisy (BAP) cases, and 41 healthy volunteers (HVs) were examined. Results: Positive range of Tm value for miR-34b/c methylation was defined as 77.71-78.79 degrees C which was the mean 3 standard deviations of 40 wells of a positive control. The number of miR-34b/c methylated wells was counted per sample according to this criterion. The number of miR-34b/c methylated wells in MPM cases was significantly higher than that in BAP cases (P = 0.03) or HVs (P < 0.001). Advanced MPM cases tended to have higher number of miR-34b/c methylated wells than early MPM cases. Receiver-operating characteristic (ROC) curve analysis revealed that three number of miR-34b/c methylated wells per sample was the best cut-off of positivity of MPM with a 67% of sensitivity and a 77% specificity for prediction. The area under the ROC curve was 0.77. Conclusions: Our digital MSP assay can quantify miR-34b/c methylation in serum-circulating DNA. The degree of miR-34b/c methylation in serum-circulating DNA is associated with MPM, suggesting that this approach might be useful for the establishment of a new detection system for MPM.
Keywords
Digital PCR
Malignant pleural mesothelioma
microRNA
miR-34b/c
Methylation
Circulating DNA
Published Date
2013-12
Publication Title
Lung Cancer
Volume
volume82
Issue
issue3
Publisher
Elsevier Ireland Ltd.
Start Page
485
End Page
490
ISSN
0169-5002
NCID
AA10785743
Content Type
Journal Article
Official Url
http://dx.doi.org/10.1016/j.lungcan.2013.09.017
Related Url
http://ousar.lib.okayama-u.ac.jp/metadata/52509
language
英語
Copyright Holders
(C) 2013 Elsevier Ireland Ltd. All rights reserved.
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Refereed
True
DOI
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