SFRP1 promotor methylation analysis of FTA card touch-prep samples derived from colonic polyps.
Adenoma
/ genetics
Adult
Aged
Aged, 80 and over
Colonic Polyps
/ genetics
DNA Methylation
/ genetics
DNA Primers
DNA, Neoplasm
/ genetics
Female
Humans
Intercellular Signaling Peptides and Proteins
/ genetics
Male
Membrane Proteins
/ genetics
Middle Aged
Polymerase Chain Reaction
Promoter Regions, Genetic
Specimen Handling
/ methods
Colorectal polyps
FTA card
Promotor methylation
Storage
Touch-prep
Journal
Experimental and molecular pathology
ISSN: 1096-0945
Titre abrégé: Exp Mol Pathol
Pays: Netherlands
ID NLM: 0370711
Informations de publication
Date de publication:
06 2020
06 2020
Historique:
received:
02
09
2019
revised:
25
01
2020
accepted:
29
01
2020
pubmed:
3
2
2020
medline:
28
10
2020
entrez:
3
2
2020
Statut:
ppublish
Résumé
Whatman FTA® cards provide the most reliable method for DNA storage and extraction, however, the literature lacks reports on the epigenetic analysis of FTA card-derived tumor DNA. Therefore, this study aimed at demonstrating that punches from colonic adenoma samples preserved on FTA filter cards are suitable for methylation analysis by real-time methylation-specific PCR (MSP). Genomic DNA was isolated from a total of 40 sporadic colorectal adenoma samples stored on FTA cards for a median of 59.60 (range 48-72) months. After bisulfite treatment, deaminated DNA was analyzed by SYBR Green real-time MSP using primers specific for methylated and unmethylated promotor sequences of the secreted frizzled-related protein 1 (SFRP1) gene. Amplifiable DNA could be isolated from all FTA card punches while SFRP1 promotor methylation was present in 34/40 (85.0%) colorectal adenomas. Our results indicate that genomic DNA isolated from colonic tumor samples preserved on FTA cards is suitable for downstream methylation detection methodologies such as MSP even after prolonged storage periods.
Identifiants
pubmed: 32007530
pii: S0014-4800(19)30676-8
doi: 10.1016/j.yexmp.2020.104397
pii:
doi:
Substances chimiques
DNA Primers
0
DNA, Neoplasm
0
Intercellular Signaling Peptides and Proteins
0
Membrane Proteins
0
SFRP1 protein, human
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
104397Informations de copyright
Copyright © 2020 Elsevier Inc. All rights reserved.