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. 2020 Oct 15;11:595902. doi: 10.3389/fmicb.2020.595902

FIGURE 4.

FIGURE 4

Loci-specific promoter methylation differences and trends among cervical cytological grades. Methylation (%) of total genomic DNA in 4 grades of cervical cytology i.e., NILM (n = 237), ASC-US (n = 235), LSIL (n = 229), and HSIL (n = 182) was compared by CpG sites among 3 genes (ADCY8, CDH8, and ZNF582). Pairwise comparisons of methylation for each CpG site between cytological grades (NILM vs. ASC-US, ASC-US vs. LSIL, and LSIL vs. HSIL) revealed significantly higher levels at multiple sites as noted by a star (* p < 0.05 by the Wilcoxon rank-sum test). Methylation levels for all CpG sites increased coincidently with cytological grade for ADCY8, CDH8, and ZNF582 by Spearman’s rs (p < 0.05, with Bonferroni adjustment) except for ADCY8 CpG sites 1–5. The methylation reference line (gray) for each assay denotes the median of the 95th percentile values for each CpG site within an assay derived from NILM (HPV-negative) samples, i.e., ADCY8 (10.11%), CDH8 (7.61%), and ZNF582 (5.22%). ASC-US, atypical squamous cells of undetermined significance; HSIL, high-grade squamous intraepithelial lesion; LSIL, low-grade squamous intraepithelial lesion; NILM, negative for intraepithelial lesion or malignancy; NS, not statistically significant.