Anti-tumor Effects and Mechanism of Recombinant Buckwheat Trypsin Inhibitor on H₂₂

Abstract: The aim of this study was to evaluate the in vitro anti-tumor effects of rBTI on hepatic cancer. Apoptosis of the H₂₂ cell line induced by rBTI was identified by MTT assays， DNA electrophoresis analysis， morphological observation of nuclei， the measurement of cytochrome c and caspases activation assess. We found that rBTI inhibited cell viability by inducing apoptosis， as evidenced by the formation of apoptotic bodies， and DNA fragmentation. rBTI induced apoptosis was correlated with mitochondrial dysfunction， leading to the release of cytochrome c from the mitochondria to the cytosol， as well as the proteolytic activation of caspase-3， caspase-8 and caspase-9. Our results suggested that one of the pathways rBTI-induced apoptosis in H₂₂ cells was mainly mediated by mitochondrial pathways via caspase-3. Moreover， rBTI could specifically inhibit the growth of cells of H₂₂ hepatic carcinoma in vitro with a concentration-dependent and time-dependent effect， but there were minimal effects on normal mice liver cell.

CSTR: 32200.14.cjcb.2009.01.0014