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Szigeti, Krisztina A; Barták, Barbara K; Nagy, Zsófia B; Zsigrai, Sára; Papp, Márton; Márkus, Eszter; Igaz, Peter; Takács, István; Molnár, Béla; Kalmár, Alexandra

Methodological and Biological Factors Influencing Global DNA Methylation Results Measured by LINE-1 Pyrosequencing Assay in Colorectal Tissue and Liquid Biopsy Samples

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  • Medientyp: E-Artikel
  • Titel: Methodological and Biological Factors Influencing Global DNA Methylation Results Measured by LINE-1 Pyrosequencing Assay in Colorectal Tissue and Liquid Biopsy Samples
  • Beteiligte: Szigeti, Krisztina A; Barták, Barbara K; Nagy, Zsófia B; Zsigrai, Sára; Papp, Márton; Márkus, Eszter; Igaz, Peter; Takács, István; Molnár, Béla; Kalmár, Alexandra
  • Erschienen: MDPI AG, 2022
  • Erschienen in: International Journal of Molecular Sciences
  • Sprache: Englisch
  • DOI: 10.3390/ijms231911608
  • ISSN: 1422-0067
  • Schlagwörter: Inorganic Chemistry ; Organic Chemistry ; Physical and Theoretical Chemistry ; Computer Science Applications ; Spectroscopy ; Molecular Biology ; General Medicine ; Catalysis
  • Entstehung:
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  • Beschreibung: <jats:p>Long interspersed nuclear element 1 (LINE-1) bisulfite pyrosequencing is a widely used technique for genome-wide methylation analyses. We aimed to investigate the effects of experimental and biological factors on its results to improve the comparability. LINE-1 bisulfite pyrosequencing was performed on colorectal tissue (n = 222), buffy coat (n = 39), and plasma samples (n = 9) of healthy individuals and patients with colorectal tumors. Significantly altered methylation was observed between investigated LINE-1 CpG positions of non-tumorous tissues (p ≤ 0.01). Formalin-fixed, paraffin-embedded biopsies (73.0 ± 5.3%) resulted in lower methylation than fresh frozen samples (76.1 ± 2.8%) (p ≤ 0.01). DNA specimens after long-term storage showed higher methylation levels (+3.2%, p ≤ 0.01). In blood collection tubes with preservatives, cfDNA and buffy coat methylation significantly changed compared to K3EDTA tubes (p ≤ 0.05). Lower methylation was detected in older (&gt;40 years, 76.8 ± 1.7%) vs. younger (78.1 ± 1.0%) female patients (p ≤ 0.05), and also in adenomatous tissues with MTHFR 677CT, or 1298AC mutations vs. wild-type (p ≤ 0.05) comparisons. Based on our findings, it is highly recommended to consider the application of standard DNA samples in the case of a possible clinical screening approach, as well as in experimental research studies.</jats:p>
  • Zugangsstatus: Freier Zugang