Twist1 was downregulated by the NF-κB–miR186–Twist1 pathway in non-transformed and normal cells (BPH1, P69), whereas this pathway was abolished due to the CpG hypermethylation of the miR186 promoter in the malignantly transformed and cancer cells (LT-BPH1, M12, PC3), thereby losing antagonization of Twist1 upregulation from the NF-κB–Twist1 pathway, which was still active. This evidence concerns the gene NFKB1 and cancer.