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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
Kishimoto, Takumi
Otsuki, Takemi
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
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