Objective: To investigate the expression of carbohydrate response element binding protein (ChREBP) and its biological actions in hepatocellular carcinoma (HCC). Methods: The mRNA and protein expressions of ChREBP in 73 specimens of HCC along with its adjacent tissue, as well as different HCC cell lines and normal hepatic cell line were determined by q-PCR, immunohistochemical staining and Western blot analysis, respectively. The changes in cell cycle, apoptosis and proliferation in HCC cells were observed after the siRNA interference of ChREBP2 expression. Results: Both mRNA and protein expressions of ChREBP were significantly increased in HCC tissue compared with tumor-adjacent tissue, and significantly increased in all studied HCC cell lines compared with normal hepatic cell line (all P<0.05). After interference of ChREBP2 expression, HCC cells presented significant G1/S phase arrest and inhibition of proliferation (both P<0.05), but apoptosis showed no significant change (P>0.05). Conclusion: ChREBP expression is elevated in HCC, which may probably promote proliferation of HCC cells through cell cycle regulation, and thereby plays an important role in the progression of HCC.