title: Anti-tumoral effects of a trypsin inhibitor derived from buckwheat in vitro and in vivo.
abstract: Native buckwheat, a common component of food products and medicine, has been observed to inhibit cancer cell proliferation in vitro. The aim of the present study was to evaluate the in vitro and in vivo anti-tumoral effects of recombinant buckwheat trypsin inhibitor (rBTI) on hepatic cancer cells and the mechanism of apoptosis involved. Apoptosis in the H22 cell line induced by rBTI was identified using MTT assays, DNA electrophoresis, flow cytometry, morphological observation of the nuclei, measurement of cytochrome C and assessment of caspase activation. It was identified that rBTI decreases cell viability by inducing apoptosis, as evidenced by the formation of apoptotic bodies and DNA fragmentation. rBTI-induced apoptosis occurred in association with mitochondrial dysfunction, leading to the release of cytochrome C from the mitochondria to the cytosol, as well as the activation of caspase-3, -8 and -9. In conclusion, the results of the present study suggested that rBTI specifically inhibited the growth of the H22 hepatic carcinoma cell line in vitro and in vivo in a concentration-dependent and time-dependent manner, while there were minimal effects on the 7702 normal liver cell line. In addition, rBTI‑induced apoptosis in H22 cells was, at least in part, mediated by a mitochondrial pathway via caspase-9.
authors.affiliation: Key Laboratory for Chemical Biology and Molecular Engineering of the Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan, Shanxi 030006, P.R. China.
authors.forename: Chong-Zhi
authors.initials: CZ
authors.lastname: Bai
authors.affiliation: Chinese Medicine Hospital of Shanxi Province, Taiyuan, Shanxi 030012, P.R. China.
authors.forename: Ma-Li
authors.initials: ML
authors.lastname: Feng
authors.affiliation: Chinese Medicine Hospital of Shanxi Province, Taiyuan, Shanxi 030012, P.R. China.
authors.forename: Xu-Liang
authors.initials: XL
authors.lastname: Hao
authors.affiliation: Key Laboratory for Chemical Biology and Molecular Engineering of the Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan, Shanxi 030006, P.R. China.
authors.forename: Zhi-Juan
authors.initials: ZJ
authors.lastname: Zhao
authors.affiliation: Key Laboratory for Chemical Biology and Molecular Engineering of the Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan, Shanxi 030006, P.R. China.
authors.forename: Yu-Ying
authors.initials: YY
authors.lastname: Li
authors.affiliation: Key Laboratory for Chemical Biology and Molecular Engineering of the Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan, Shanxi 030006, P.R. China.
authors.forename: Zhuan-Hua
authors.initials: ZH
authors.lastname: Wang
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D019612:Fagopyrum / Q000737:chemistry* / Q000378:metabolism; D005260:Female; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D014183:Transplantation, Heterologous; D014361:Trypsin Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use
chemical_list: D000970:Antineoplastic Agents; D014361:Trypsin Inhibitors; D045304:Cytochromes c; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9