Abstract: Objective To screen differentially expressed genes in cytochalasin B (CB)-induced denucleated K562 cells by restriction display (RD) technique. Methods The total RNA was isolated and purified from K562 cells before and after CB (10 μg/ml) treatment. The mRNA from both treated and untreated K562 cells were reversly transcribed into cDNA, and the differentially expressed genes were separated using RD technique combined with polyacrylamide gel electrophoresis and sliver staining, followed by cloning, sequencing and homology analysis against GenBank database of these genes. Results Seven differentially expressed genes were identified in CB-treated cells including aquaporin 1 (AQP1) gene, which was verified to be up-regulated after CB treatment by RT-PCR. Conclusion AQP1 gene might be in close association with the regulation of de-nucleation processes and CB-induced proliferation inhibition of K562 cells.