Herein, we provide a comprehensive overview of the fast-evolving field of pharmacological targeting of signaling pathways by thymoquinone in cancer and discuss the possibilities associated with targeting JAK/STAT, Wnt/β-catenin, PI3K/AKT/mTOR, NF-κB, and TRAIL-driven pathways for drug development in a wide range of oncology settings. This evidence concerns the gene NFKB1 and cancer.