Standardization of DNA amount for bisulfite conversion for analyzing the methylation status of LINE-1 in lung cancer.

Highly methylated Long Interspersed Nucleotide Elements 1 (LINE-1) constitute approximately 20% of the human genome, thus serving as a surrogate marker of global genomic DNA methylation. To date, there is still lacking a consensus about the precise location in LINE-1 promoter and its methylation thr...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Duong Anh Thuy Pham, Son Duc Le, Trang Mai Doan, Phuong Thu Luu, Uyen Quynh Nguyen, Son Van Ho, Lan Thi Thuong Vo
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a021c701ba8c431cbd606617988a746b
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:a021c701ba8c431cbd606617988a746b
record_format	dspace
spelling	oai:doaj.org-article:a021c701ba8c431cbd606617988a746b2021-12-02T20:17:54ZStandardization of DNA amount for bisulfite conversion for analyzing the methylation status of LINE-1 in lung cancer.1932-620310.1371/journal.pone.0256254https://doaj.org/article/a021c701ba8c431cbd606617988a746b2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0256254https://doaj.org/toc/1932-6203Highly methylated Long Interspersed Nucleotide Elements 1 (LINE-1) constitute approximately 20% of the human genome, thus serving as a surrogate marker of global genomic DNA methylation. To date, there is still lacking a consensus about the precise location in LINE-1 promoter and its methylation threshold value, making challenging the use of LINE-1 methylation as a diagnostic, prognostic markers in cancer. This study reports on a technical standardization of bisulfite-based DNA methylation analysis, which ensures the complete bisulfite conversion of repeated LINE-1 sequences, thus allowing accurate LINE-1 methylation value. In addition, the study also indicated the precise location in LINE-1 promoter of which significant variance in methylation level makes LINE-1 methylation as a potential diagnostic biomarker for lung cancer. A serial concentration of 5-50-500 ng of DNA from 275 formalin-fixed paraffin-embedded lung tissues were converted by bisulfite; methylation level of two local regions (at nucleotide position 300-368 as LINE-1.1 and 368-460 as LINE-1.2) in LINE-1 promoter was measured by real time PCR. The use of 5 ng of genomic DNA but no more allowed to detect LINE-1 hypomethylation in lung cancer tissue (14.34% versus 16.69% in non-cancerous lung diseases for LINE-1.1, p < 0.0001, and 30.28% versus 32.35% for LINE-1.2, p < 0.05). Our study thus highlighted the optimal and primordial concentration less than 5 ng of genomic DNA guarantees the complete LINE-1 bisulfite conversion, and significant variance in methylation level of the LINE-1 sequence position from 300 to 368 allowed to discriminate lung cancer from non-cancer samples.Duong Anh Thuy PhamSon Duc LeTrang Mai DoanPhuong Thu LuuUyen Quynh NguyenSon Van HoLan Thi Thuong VoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 8, p e0256254 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Duong Anh Thuy Pham Son Duc Le Trang Mai Doan Phuong Thu Luu Uyen Quynh Nguyen Son Van Ho Lan Thi Thuong Vo Standardization of DNA amount for bisulfite conversion for analyzing the methylation status of LINE-1 in lung cancer.
description	Highly methylated Long Interspersed Nucleotide Elements 1 (LINE-1) constitute approximately 20% of the human genome, thus serving as a surrogate marker of global genomic DNA methylation. To date, there is still lacking a consensus about the precise location in LINE-1 promoter and its methylation threshold value, making challenging the use of LINE-1 methylation as a diagnostic, prognostic markers in cancer. This study reports on a technical standardization of bisulfite-based DNA methylation analysis, which ensures the complete bisulfite conversion of repeated LINE-1 sequences, thus allowing accurate LINE-1 methylation value. In addition, the study also indicated the precise location in LINE-1 promoter of which significant variance in methylation level makes LINE-1 methylation as a potential diagnostic biomarker for lung cancer. A serial concentration of 5-50-500 ng of DNA from 275 formalin-fixed paraffin-embedded lung tissues were converted by bisulfite; methylation level of two local regions (at nucleotide position 300-368 as LINE-1.1 and 368-460 as LINE-1.2) in LINE-1 promoter was measured by real time PCR. The use of 5 ng of genomic DNA but no more allowed to detect LINE-1 hypomethylation in lung cancer tissue (14.34% versus 16.69% in non-cancerous lung diseases for LINE-1.1, p < 0.0001, and 30.28% versus 32.35% for LINE-1.2, p < 0.05). Our study thus highlighted the optimal and primordial concentration less than 5 ng of genomic DNA guarantees the complete LINE-1 bisulfite conversion, and significant variance in methylation level of the LINE-1 sequence position from 300 to 368 allowed to discriminate lung cancer from non-cancer samples.
format	article
author	Duong Anh Thuy Pham Son Duc Le Trang Mai Doan Phuong Thu Luu Uyen Quynh Nguyen Son Van Ho Lan Thi Thuong Vo
author_facet	Duong Anh Thuy Pham Son Duc Le Trang Mai Doan Phuong Thu Luu Uyen Quynh Nguyen Son Van Ho Lan Thi Thuong Vo
author_sort	Duong Anh Thuy Pham
title	Standardization of DNA amount for bisulfite conversion for analyzing the methylation status of LINE-1 in lung cancer.
title_short	Standardization of DNA amount for bisulfite conversion for analyzing the methylation status of LINE-1 in lung cancer.
title_full	Standardization of DNA amount for bisulfite conversion for analyzing the methylation status of LINE-1 in lung cancer.
title_fullStr	Standardization of DNA amount for bisulfite conversion for analyzing the methylation status of LINE-1 in lung cancer.
title_full_unstemmed	Standardization of DNA amount for bisulfite conversion for analyzing the methylation status of LINE-1 in lung cancer.
title_sort	standardization of dna amount for bisulfite conversion for analyzing the methylation status of line-1 in lung cancer.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/a021c701ba8c431cbd606617988a746b
work_keys_str_mv	AT duonganhthuypham standardizationofdnaamountforbisulfiteconversionforanalyzingthemethylationstatusofline1inlungcancer AT sonducle standardizationofdnaamountforbisulfiteconversionforanalyzingthemethylationstatusofline1inlungcancer AT trangmaidoan standardizationofdnaamountforbisulfiteconversionforanalyzingthemethylationstatusofline1inlungcancer AT phuongthuluu standardizationofdnaamountforbisulfiteconversionforanalyzingthemethylationstatusofline1inlungcancer AT uyenquynhnguyen standardizationofdnaamountforbisulfiteconversionforanalyzingthemethylationstatusofline1inlungcancer AT sonvanho standardizationofdnaamountforbisulfiteconversionforanalyzingthemethylationstatusofline1inlungcancer AT lanthithuongvo standardizationofdnaamountforbisulfiteconversionforanalyzingthemethylationstatusofline1inlungcancer
_version_	1718374362426376192

Standardization of DNA amount for bisulfite conversion for analyzing the methylation status of LINE-1 in lung cancer.

Ejemplares similares