The qMSP results confirmed significant hypomethylation of EN1 in SACC tissues compared with paired normal tissues (R2 = 0.9644) (Fig. 6C), with a statistically significant difference in the overall methylation level between the tumour and normal tissues (median PMR, 0.34 vs. 0.56, P = 0.0027) (Fig. 6D). This evidence concerns the gene EN1 and neoplasm.