Anti-tumoral effects of a trypsin inhibitor derived from buckwheat in vitro and in vivo

Bai,Chong‑Zhi; Feng,Ma‑Li; Hao,Xu‑Liang; Zhao,Zhi‑Juan; Li,Yu‑Ying; Wang,Zhuan‑Hua

doi:10.3892/mmr.2015.3649

Anti-tumoral effects of a trypsin inhibitor derived from buckwheat in vitro and in vivo

Authors:
- Chong‑Zhi Bai
- Ma‑Li Feng
- Xu‑Liang Hao
- Zhi‑Juan Zhao
- Yu‑Ying Li
- Zhuan‑Hua Wang
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Affiliations: Key Laboratory for Chemical Biology and Molecular Engineering of the Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan, Shanxi 030006, P.R. China, Chinese Medicine Hospital of Shanxi Province, Taiyuan, Shanxi 030012, P.R. China
Published online on: April 20, 2015 https://doi.org/10.3892/mmr.2015.3649
Pages: 1777-1782
Copyright: © Bai et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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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.

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