Objective To explore the interaction between LIM protein Ajuba and transcription factor Twist in colorectal cancer cells and the effects of this interaction on the migration ability of colorectal cancer cells. Methods Co-immunoprecipitation test was used to verify whether the interaction exists between exogenous Ajuba and Twist. The transcription activity of Ajuba towards N-cadherin promoter of downstream target gene of Twist was detected by the luciferase activity assay. Ajuba knock-down stable cell lines SW1116-shAjuba were established. Changes of the cell migration ability were detected by the Transwell assay. Results The interaction between Ajuba and Twist existed and Ajuba could enhance the regulation of N-cadherin expression through Twist. The expression of Ajuba gene in SW1116-shAjuba cells decreased; protein expression down-regulated; and cell migration ability was inhibited. Conclusion Ajuba is a new coactivator of transcription factor Twist. Ajuba can promote the expression of N-cadherin and enhance the migration ability of colorectal cancer cells.