Objective:Zinc-alpha-2-glycoprotein 1(ZAG) is a protein with multiple functions,such as participating in lipometabolism,saccharometabolism,energy metabolism,immunoreaction and fertilization. It is observed whether ZAG involved in the regulation of human colorectal cancer cell line(HT-29) in this study. Methods:ZAG-RNAi plasmids were constructed and transfected into human colorectal cancer HT-29 cells. The expressions of target proteins and gene were detected by Western blot and RT-PCR,respectively. Adenosine triphosphate (ATP) was assessed to investigate the activation of energy metabolism. The capacity of HT-29 cell migration was investigated using the Transwell migration assay. Results:ZAG interference plasmids were successfully transfected into HT-29 cells,and stably transfected cell lines were screened out. ATP level and the migration of ZAG inhibitor-treated cells were increased. Conclusion:The down-expression of ZAG promoted the expression of ATP and up-regulated the malignant phenotype of HT-29 cells.