To further evaluate the prognostic significance of these two m5C-related regulators, LASSO Cox regression analysis was performed and it was revealed that NSUN6 (Coef = 0.300256795278519) and ALYREF (Coef = 0.00796895949684636) could serve as powerful prognostic factors in COAD (Figures 7B–C). This evidence concerns the gene ALYREF and colon adenocarcinoma.