The role of Wnt/β-catenin signaling pathway in the etiology and/or progression of ovarian cancer has been well documented. It was demonstrated that ovarian cancer constantly exhibit constitutive activation of canonical Wnt signaling, usually as a result of oncogenic mutations that stabilize and dysregulate the β-catenin protein. In this study, we transfected an expression vector-based small hairpin RNA (shRNA) targeting to β-catenin encoding gene into human A2780 ovarian cancer cells to investigate the effects of β-catenin knockdown on biological characteristics of ovarian cancer cells. The results showed that β-catenin shRNA expression resulted in decreased β-catenin mRNA and protein expression in the transfected A2780 cells, inhibition of cellular proliferation, decreased capability of clonogenicity in the plating and the soft agar, and increased sensitivities to chemotherapy drugs vincristine, paclitaxel and cisplatin compared to untransfected cells. Importantly, we found that shRNA-mediated knockdown of β-catenin strongly decreases tumour growth of human A2780 ovarian cancer cells in xenografts. These results demonstrate that β-catenin might be an effective therapeutic target for human ovarian cancer treatment.