Effect of Recombinant Buckwheat Trypsin Inhibitor on Apoptosis and
Caspases Activity in Human Hepatoma (HepG2) Cells

LIFang;LIYu-Ying;BAIChong-Zhi;TIANXin;ZHANGZheng;WANGZhuan-Hua

Chinese Journal of Biochemistry and Molecular Biology ›› 2009, Vol. 25 ›› Issue (02) : 182-187.

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PDF(365 KB)
Chinese Journal of Biochemistry and Molecular Biology ›› 2009, Vol. 25 ›› Issue (02) : 182-187.
Research Papers

Effect of Recombinant Buckwheat Trypsin Inhibitor on Apoptosis and
Caspases Activity in Human Hepatoma (HepG2) Cells

  • LI Fang;LI Yu-Ying;BAI Chong-Zhi;TIAN Xin;ZHANG Zheng;WANG Zhuan-Hua*

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Abstract

Recombinant buckwheat trypsin inhibitor (rBTI) has been reported to induce apoptosis in human cancer cells,such as IM-9,K562.To examine whether rBTI had an anticancer effect on HepG2 human hepatoma cells,and reveal the possible mechanisms,cells treated with rBTI were subjected to MTT,nuclear staining and DNA fragmentation assays for measuring growth inhibition and induction of apoptosis.In addition,as the Bcl-2 (an anti-apoptotic gene)/Bax (a pro-apoptotic gene) ratio is associated with the increase of apoptosis,the mRNA expression levels of both were determined.Following rBTI incubation,Bcl-2 was decreased,whereas Bax was increased.We also investigated the alteration of the Fas/FasL system after rBTI treatment in HepG2 cells. However,not significant changes were found.rBTI treatments markedly induced the activities of caspase-3 and caspase-9,while caspase-8 was responded slightly.These findings suggested that rBTIinduced apoptosis was likely initiated at mitochondria and maybe related to the caspase-3 signaling pathway,but not via Fas/FasL.

Key words

recombinant buckwheat trypsin inhibitor / apoptosis / HepG2 cell / caspases

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LIFang;LIYu-Ying;BAIChong-Zhi;TIANXin;ZHANGZheng;WANGZhuan-Hua.

Effect of Recombinant Buckwheat Trypsin Inhibitor on Apoptosis and
Caspases Activity in Human Hepatoma (HepG2) Cells

[J]. Chinese Journal of Biochemistry and Molecular Biology, 2009, 25(02): 182-187

References

[1] Foehr M W, Tomei L D, Goddard J G, et al. Antiapoptotic activity of the Bowman-Birk inhibitor can be attributed to copurified phospholipids [J]. Nutr Cancer, 1999, 34 (2): 199-205
[2] Inoue K, Takano H, Yanagisawa R, et al. Protective role of urinary trypsin inhibitor in acute lung injury induced by lipopolysaccharide [J]. Exp Biol Med (Maywood), 2005, 230 (4): 281-287
[3] 李丽琴, 郑晓军, 陈乐贵, 等. 蛋白酶体抑制剂药理活性的研究进展[J]. 国外医学药学分册 (Li Li-Qing, Zheng Xiao-Jun, Chen Le-Gui, et al. The research progress on proteinase inhibitors pharmacological activity [J]. Foreign Med Sci-Section Pharm), 2001, 28 (3): 132-136
[4] Gladysheva I P, Moroz N A, Karmakova T A, et al. Immunoconjugates of soybean Bowman-Birk protease inhibitor as targeted antitumor polymeric agents [J]. J Drug Target, 2001, 9 (5): 303-316
[5] Malkowicz S B, Liu S P, Broderick G A, et al. Effect of the Bowman-Birk inhibitor (a Soy Protein) on in vitro bladder neck/urethral and penile corporal smooth muscle activity[J]. Neurourol Urodyn, 2003, 22 (1): 54-57
[6] Ren W, Qiao Z, Wang H, et al. Molecular basis of Fas and cytochrome c pathways of apoptosis induced by tartary buckwheat flavonoid in HL-60 cells [J]. Methods Find Exp Clin Pharmacol, 2003, 25 (6):431-436
[7] 高丽, 李玉英, 张政,等. 重组荞麦胰蛋白酶抑制剂对HL-60细胞的促凋亡作用[J]. 中国实验血液
学杂志( Gao Li, Li Yu-Ying, Zhang Zheng, et al. Apoptosis of human leukemia HL-60 cells induced by recombinant common buckwheat trypsin inhibitor [J]. J Exp Hematol), 2007, 15 (1):59-62
[8] Zhang Z, Li Y Y, Li C, et al. Expression of a buckwheat trypsin inhibitor in Escherichia coli and its
effect on proliferation of multiple myeloma IM-9 cell proliferation[J]. Acta Biochim Biophys Sin, 2007,
39 (9): 701-707
[9] Wang Z H, Gao L, Li Y Y, et al. Induction of apoptosis by buckwheat trypsin inhibitor in chronic
myeloid leukemia K562 cells [J]. Biol Pharm Bull, 2007, 30 (4): 783-786
[10] Li J, Xia X, Ke Y, et al. Trichosanthin induced apoptosis in HL-60 cells via mitochondrial and endoplasmic reticulum stress signaling pathways [J]. Biochim Biophys Acta, 2007, 1770 (8): 1169-1180
[11] 韩雪莲,方志杰,陈军浩,等. 合成芪类化合物抑制肿瘤细胞增殖并诱导Bel-7402 细胞凋亡
[J].中国生物化学与分子生物学报(Han Xue-Lian, Fang Zhi-Jie, Chen Jun-Hao, et al. Stilbene compounds inhibit proliferation of tumor cells and induce apoptosis of Bel-7402 cells[J]. Chin J Biochem Mol Biol), 2007, 23(11):963-970
[12] 刘权焰, 刘志苏, 邬开朗, 等. MAT2A基因小干扰RNA诱导人肝癌细胞凋亡的分子机制[J]. 中国生物化学与分子生物学报 (Liu Quan-Yan, Liu Zhi-Su, Wu Kai-Lang, et al. Silencing of MAT2A gene by RNA interference resulted in induction of apoptosis in human hepatoma cells[J]. Chin J Biochem Mol Biol), 2006,22 (7): 575-583
[13] 刘闻闻,于春晓,崔福爱,等. 人同源盒基因NKX3.1对前列腺癌细胞的诱导凋亡作用[J]. 中国
生物化学与分子生物学报 (Liu Wen-Wen,Yu Chun-Xiao,Cui Fu-Ai, et al. Apoptosis-inducing effect of human homeobox gene NKX3.1 on prostate cancer cells [J].Chin J Biochem Mol Biol),2006, 22 (12): 996-1002
[14]王多宁, 赵雁武, 田芙蓉,等. 考马斯亮蓝微盘比色法测定蛋白质含量[J]. 第四军医大学学报(Wang Duo-Ning, Zhao Yan-Wu, Tian Fu-Rong,et al. Protein quantification with Coomassie bulliant blue microplate-colormetric [J]. J Fourth Mil Med Univ), 2001, 22 (6): 528-529
[15] Jin W, Di G, Li J, et al. TIEG1 induces apoptosis through mitochondrial apoptotic pathway and promotes apoptosis induced by homoharringtonine and velcade [J]. FEBS Lett, 2007, 581 (20): 3826-3832
[16] Lei X, Chen Y, Du G, et al. Gossypol induces Bax/Bak-independent activation of apoptosis and cytochrome c release via a conformational change in Bcl-2 [J]. FASEB J, 2006, 20(12): 2147-2149
[17] 刘志, 郑军. Bcl-2家族蛋白及其在细胞凋亡中的作用[J]. 生命的化学( Liu Zhi, Zheng Jun. Bcl-2 family proteins and the role in apoptosis [J]. Chem Life ), 2007, 27 (1): 22-25
[18] 金洪传. Fas介导的细胞凋亡途径[J].浙江大学学报(医学版)(Jin Hong-Chuan. Fas-mediated apoptosis pathway [J]. J Zhejiang Univ (Med Sci) ), 1999, 28 (5): 237-240
[19] Kim M J, Kim Y J, Park H J, et al. Apoptotic effect of red wine polyphenols on human colon cancer SNU-C4 cells [J]. Food Chem Toxicol, 2006, 44 (6): 898-902
[20] Woo H J, Lee S J, Choi B T, et al. Induction of apoptosis and inhibition of telomerase activity by trichostatin A, a histone deacetylase inhibitor, in human leukemic U937 cells [J]. Exp Mol Pathol, 2007, 82 (1): 77-84
[21] Zhang L, Cui L. A cytotoxin isolated from Agkistrodon acutus snake venom induces apoptosis via Fas pathway in A549 cells [J]. Toxicol In Vitro, 2007, 21 (6):1095-1103
[22] Raisova M, Bektas M, Wieder T, et al. Resistance to CD95/Fas-induced and ceramide-mediated apoptosis of human melanoma cells is caused by a defective mitochondrial cytochrome c release [J]. FEBS Lett, 2000, 473 (1): 27-32
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