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Zhao-Xiong Zhang,1 Wan-Ni Zhang,1 Yuan-Yuan Sun,1 Yun-Hui Li,2 Zhen-Ming Xu,3 Wei-Neng Fu1 1Department of Medical Genetics, China Medical University, Shenyang, People’s Republic of China; 2Department of Laboratory Medicine, No 202 Hospital of PLA, Shenyang, People’s Republic of China; 3Department of Otolaryngology, No 463 Hospital of PLA, Shenyang, People’s Republic of China Purpose: CREB, MYCY1 and NAT10 are involved in cancer cell migration. However, the relationship between these three proteins and their role in laryngeal cancer cell migration remains unknown. Methods: Transient gene transfection was performed in laryngeal cancer cells. Bioinformatics analysis was used to predict the binding of CREB to MYCT1 promoter. Binding of CREB to the promoter of MYCT1 was monitored by luciferase reporter assay and chromatin immunoprecipitation method in vitro and in vivo, respectively. Real-time RT-PCR and Western bolt were applied to detect gene transcription and translation levels, respectively. Laryngeal cancer cell migration was assayed by transwell cham­ber experiment. Results: CREB protein expression was significantly up-regulated in laryngeal cancer tissues and associated with cancer differentiation, tumor stage, and lymphatic metasta­sis. CREB inhibits MYCT1 expression by direct binding to its promoter. Meanwhile, MYCT1 has a negative impact on the NAT10 gene expression. Furthermore, CREB promotes NAT10 expression via down-regulating the MYCT1 gene expression. In addition, contrary to MYCT1, CREB and NAT10 enhanced laryngeal cancer cell migration. MYCT1 and NAT10 significantly rescued the effects of CREB and MYCT1 on Hep2 cell migration, respectively. Conclusion: CREB promotes laryngeal cancer cell migration via MYCT1/NAT10 axis, suggesting that CREB might be a potential prognostic marker in laryngeal cancer. Keywords: laryngeal cancer, CREB, MYCT1, NAT10, migration