Detection of MSH2 Gene Methylation in Extramammary Paget’s Disease by Methylation-Sensitive High-Resolution Melting Analysis

Background. Extramammary Paget’s disease (EMPD) is a rare skin tumor. Hypermethylation in the MSH2 promoter resulting in the downregulation of its protein expression shows a high detection rate in EMPD tumor tissue, which indicates that the methylation of MSH2 may play an important role in the patho...

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Autores principales:	Liu Dong, Yingfeng Zhu, Liting Wu, Qiaoan Zhang, Feng Xu, Xinju Zhang, Xiao Xu, Yiting Tang, Guoqiang Ren, Zhihua Kang, Ming Guan
Formato:	article
Lenguaje:	EN
Publicado:	Hindawi Limited 2021
Materias:	Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282
Acceso en línea:	https://doaj.org/article/0c12d2a225ad40feaec04452c394e83b
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spelling	oai:doaj.org-article:0c12d2a225ad40feaec04452c394e83b2021-11-15T01:19:55ZDetection of MSH2 Gene Methylation in Extramammary Paget’s Disease by Methylation-Sensitive High-Resolution Melting Analysis1687-846910.1155/2021/5514426https://doaj.org/article/0c12d2a225ad40feaec04452c394e83b2021-01-01T00:00:00Zhttp://dx.doi.org/10.1155/2021/5514426https://doaj.org/toc/1687-8469Background. Extramammary Paget’s disease (EMPD) is a rare skin tumor. Hypermethylation in the MSH2 promoter resulting in the downregulation of its protein expression shows a high detection rate in EMPD tumor tissue, which indicates that the methylation of MSH2 may play an important role in the pathogenesis of EMPD. Objective. This study aims to establish a rapid analysis strategy based on the methylation-sensitive high-resolution melting curve (MS-HRM) to detect the methylation level of the MSH2 promoter. Methods. With the use of universal methylated human DNA products, we established the MS-HRM standard curve to quantitatively detect the methylation level of the MSH2 promoter. Then, all 57 EMPD tumor DNA samples were analyzed. Pyrosequencing assay was also carried out to test the accuracy and efficacy of MS-HRM. Besides, a total of 54 human normal and other cancerous tissues were included in this study to test the reliability and versatility of the MS-HRM standard curve. Results. In this study, by using the established MS-HRM, we found that 96.5% (55/57) EMPD tumor samples had varying methylation levels in the MSH2 promoter ranging from 0% to 30%. Then, the methylation data were compared to the results obtained from pyrosequencing, which showed a high correlation between these two techniques by Pearson’s correlation (r = 0.9425) and Bland–Altman plots (mean difference = −0.1069) indicating that the methylation levels analyzed by MS-HRM were consistent with DNA pyrosequencing. Furthermore, in 23 normal and 31 other cancerous tissue samples, there were two colorectal cancer (CRC) tissues that tested MSH2 methylation positive (1% and 5%) which confirmed that our established MS-HRM can be widely applied to various types of samples. Conclusion. MS-HRM standard curve can be used for the detection of the methylation level of MSH2 in EMPD tumor samples and other cancerous tissues potentially, which presents a promising candidate as a quantitative assay to analyze MSH2 promoter methylation in routine pathological procedure.Liu DongYingfeng ZhuLiting WuQiaoan ZhangFeng XuXinju ZhangXiao XuYiting TangGuoqiang RenZhihua KangMing GuanHindawi LimitedarticleNeoplasms. Tumors. Oncology. Including cancer and carcinogensRC254-282ENJournal of Oncology, Vol 2021 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282
spellingShingle	Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282 Liu Dong Yingfeng Zhu Liting Wu Qiaoan Zhang Feng Xu Xinju Zhang Xiao Xu Yiting Tang Guoqiang Ren Zhihua Kang Ming Guan Detection of MSH2 Gene Methylation in Extramammary Paget’s Disease by Methylation-Sensitive High-Resolution Melting Analysis
description	Background. Extramammary Paget’s disease (EMPD) is a rare skin tumor. Hypermethylation in the MSH2 promoter resulting in the downregulation of its protein expression shows a high detection rate in EMPD tumor tissue, which indicates that the methylation of MSH2 may play an important role in the pathogenesis of EMPD. Objective. This study aims to establish a rapid analysis strategy based on the methylation-sensitive high-resolution melting curve (MS-HRM) to detect the methylation level of the MSH2 promoter. Methods. With the use of universal methylated human DNA products, we established the MS-HRM standard curve to quantitatively detect the methylation level of the MSH2 promoter. Then, all 57 EMPD tumor DNA samples were analyzed. Pyrosequencing assay was also carried out to test the accuracy and efficacy of MS-HRM. Besides, a total of 54 human normal and other cancerous tissues were included in this study to test the reliability and versatility of the MS-HRM standard curve. Results. In this study, by using the established MS-HRM, we found that 96.5% (55/57) EMPD tumor samples had varying methylation levels in the MSH2 promoter ranging from 0% to 30%. Then, the methylation data were compared to the results obtained from pyrosequencing, which showed a high correlation between these two techniques by Pearson’s correlation (r = 0.9425) and Bland–Altman plots (mean difference = −0.1069) indicating that the methylation levels analyzed by MS-HRM were consistent with DNA pyrosequencing. Furthermore, in 23 normal and 31 other cancerous tissue samples, there were two colorectal cancer (CRC) tissues that tested MSH2 methylation positive (1% and 5%) which confirmed that our established MS-HRM can be widely applied to various types of samples. Conclusion. MS-HRM standard curve can be used for the detection of the methylation level of MSH2 in EMPD tumor samples and other cancerous tissues potentially, which presents a promising candidate as a quantitative assay to analyze MSH2 promoter methylation in routine pathological procedure.
format	article
author	Liu Dong Yingfeng Zhu Liting Wu Qiaoan Zhang Feng Xu Xinju Zhang Xiao Xu Yiting Tang Guoqiang Ren Zhihua Kang Ming Guan
author_facet	Liu Dong Yingfeng Zhu Liting Wu Qiaoan Zhang Feng Xu Xinju Zhang Xiao Xu Yiting Tang Guoqiang Ren Zhihua Kang Ming Guan
author_sort	Liu Dong
title	Detection of MSH2 Gene Methylation in Extramammary Paget’s Disease by Methylation-Sensitive High-Resolution Melting Analysis
title_short	Detection of MSH2 Gene Methylation in Extramammary Paget’s Disease by Methylation-Sensitive High-Resolution Melting Analysis
title_full	Detection of MSH2 Gene Methylation in Extramammary Paget’s Disease by Methylation-Sensitive High-Resolution Melting Analysis
title_fullStr	Detection of MSH2 Gene Methylation in Extramammary Paget’s Disease by Methylation-Sensitive High-Resolution Melting Analysis
title_full_unstemmed	Detection of MSH2 Gene Methylation in Extramammary Paget’s Disease by Methylation-Sensitive High-Resolution Melting Analysis
title_sort	detection of msh2 gene methylation in extramammary paget’s disease by methylation-sensitive high-resolution melting analysis
publisher	Hindawi Limited
publishDate	2021
url	https://doaj.org/article/0c12d2a225ad40feaec04452c394e83b
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Detection of MSH2 Gene Methylation in Extramammary Paget’s Disease by Methylation-Sensitive High-Resolution Melting Analysis

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