Abstract βII-spectrin(SPTBN1, β2SP), the most common nonerythrocytic member of the β-spectrin gene family, functions as an adapter protein for Smad3/Smad4 complex formation during TGF-β signal transduction, and is required for EMT during embryonic liver development. Moreover, β2SP is emerging as a tumor suppressor. We have reported that Loss of β2SP enhances the stem-like traits and invasiveness of HCC cells. The canonical Wnt pathway, known to be a critical regulator of self-renewal in stem cell, is also constitutively activated and implicated in the induction of EMT in cancer. Kallistatin (a serine protease inhibitor, SERPINA4), which was identified as a unique inhibitor of the Wnt pathway, is expressed primly in the liver. In this study, we investigated the mechanism by which β2SP influences stem cell traits and aggressive behavior of HCC cell lines. We found that cells with decreased β2SP have decreased β-catenin phosphorylation and increased β-catenin nuclear localization, indicative of activation of Wnt signaling. Furthermore, loss of β2SP results in the decease of Kallistatin expression, the Wnt inhibitor. Restore of kallistatin reversed the Wnt activation in HCC cells from decreased β2SP level. Our study provides evidence for the first time that loss of β2SP activates Wnt signal through suppressing kallistatin expression. Note: This abstract was not presented at the meeting. Citation Format: Xiuling Zhi, Ling Lin, Narayan Shivapurkar Shivapurkar, John L. Marshall, Lynt Johnson, Aiwu R. He. βII-spectrin(SPTBN1) suppresses tumor progression by inhibiting Wnt signaling via kallistatin in hepatocellular carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5275. doi:10.1158/1538-7445.AM2014-5275