Thereafter, based on the expression of the m5C-related regulators, patients with COAD were clustered into two subgroups (cluster I and cluster II), an the cluster II had a longer survival time than cluster I. To further study the effect of m5C-related regulators on the prognosis and clinicopathological characteristics of COAD, we constructed a prognostic risk signature using two identified m5C-related regulators (NSUN6 and ALYREF) and were able to assign patients with COAD into high- and low-risk groups. This evidence concerns the gene NSUN6 and colon adenocarcinoma.