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. 2020 Jun 28;21(13):4595. doi: 10.3390/ijms21134595

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Developments of CDH1/CDKN2Ap16/RASSF1A/TERT/WT1 methylation levels. (A) Average promoter methylation levels for each gene of interest for tumor-free control samples (n_CDH1 = 2; n_CDKN2Ap16 = 7; n_RASSF1A = 1; n_TERT = 7; n_WT1 = 3), wildtype (n_CDH1 = 34; n_CDKN2Ap16 = 89; n_RASSF1A = 22; n_TERT = 78; n_WT1 = 23) and EGFR/KRAS mutated tumor samples (n_CDH1 = 22; n_CDKN2Ap16 = 46; n_RASSF1A = 19; n_TERT = 39; n_WT1 = 16). Significant changes are measured for TERT methylation; i.e., G0 vs. mutated, p = 0.050 and mutated vs. wild-type p = 0.046. Student’s t-test. (B) Average methylation of each gene in non-tumor/G0, low grade (G1-G2) (n_CDH1 = 14; n_CDKN2Ap16 = 43; n_RASSF1A = 11; n_TERT = 33; n_WT1 = 18), and high grade tumor samples (G3-G4) (n_CDH1 = 42; n_CDKN2Ap16 = 92; n_RASSF1A = 30; n_TERT = 85; n_WT1 = 21). A statistically significant increase of TERT methylation was found comparing tumor-free control to low-grade tumors (p = 0.022) and to high grade tumors (p = 0.021), the steady increase with tumor progression is a significant correlation (p = 0.003) Spearman correlation. WT1 methylation decreased significantly comparing tumor-free control to low grade tumor samples (p = 0.043) Student’s t-test. (C) Methylation of target genes in the patient samples grouped by gender and age. A Spearman correlation analysis matrix showed no significant correlation between overall age and promoter methylation for any of the analyzed genes. * p < 0.05.