The type-I ribosome-inactivating protein trichosanthin (TCS) has a broad spectrum of biological and pharmacological activities, including abortifacient, anti-tumour and anti-HIV activities. We have found for the first time that TCS stimulated the production of reactive oxygen species (ROS) in JAR cells (a human choriocarcinoma cell line) in a time- and concentration-dependent manner by using the fluorescent probe 2′,7′-dichlorofluorescein diacetate with confocal laser scanning microscopy. ESR spectral studies and the inhibition of ROS formation by the superoxide radical anion (O2-P) scavenger superoxide dismutase, the H2O2 scavenger catalase and the hydroxyl radical (OHP) scavenger mannitol suggested the involvement of O2-P, H2O2 and OHP. TCS-induced ROS formation was shown to be dependent on the presence of both extracellular and intracellular Ca2+; moreover, ROS production paralleled the intracellular Ca2+ elevation induced by TCS, suggesting that ROS production might be a consequence of Ca2+ signalling. TCS-induced activation of caspase-3 was initiated within 2h; however, TCS-induced production of ROS was initiated within 5min, suggesting that the production of ROS preceded the activation of caspase-3. Simultaneous observation of the nuclear morphological changes via two-photon laser scanning microscopy and ROS production via confocal laser scanning microscopy revealed that ROS is involved in the apoptosis of JAR cells. The involvement of ROS was also confirmed by the inhibition of TCS-induced cell death by the antioxidant Trolox and the ROS scavengers catalase and mannitol. Diethylenetriaminepenta-acetic acid, an inhibitor of metal-facilitated OHP formation, markedly inhibited TCS-induced cell death, suggesting that TCS induced OHP formation via the Fenton reaction. The finding that ROS is involved in the TCS-induced apoptosis of JAR cells might provide new insight into the anti-tumour and anti-HIV mechanism of TCS.
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May 2001
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Research Article|
April 24 2001
Reactive oxygen species involved in trichosanthin-induced apoptosis of human choriocarcinoma cells
Chun-yang ZHANG;
Chun-yang ZHANG
*Molecular and Nano Sciences Laboratory, Department of Physics, Tsinghua University, Beijing 100084, China
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Yi-xuan GONG;
Yi-xuan GONG
†National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China
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Hui MA;
Hui MA
1
*Molecular and Nano Sciences Laboratory, Department of Physics, Tsinghua University, Beijing 100084, China
1To whom correspondence should be addressed (e-mail [email protected]).
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Cheng-cai AN;
Cheng-cai AN
†National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China
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Die-yan CHEN
Die-yan CHEN
*Molecular and Nano Sciences Laboratory, Department of Physics, Tsinghua University, Beijing 100084, China
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Publisher: Portland Press Ltd
Received:
October 30 2000
Revision Received:
January 18 2001
Accepted:
February 13 2001
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 2001
2001
Biochem J (2001) 355 (3): 653–661.
Article history
Received:
October 30 2000
Revision Received:
January 18 2001
Accepted:
February 13 2001
Connected Content
A correction has been published:
Reactive oxygen species involved in trichosanthin-induced apoptosis of human choriocarcinoma cells
Citation
Chun-yang ZHANG, Yi-xuan GONG, Hui MA, Cheng-cai AN, Die-yan CHEN; Reactive oxygen species involved in trichosanthin-induced apoptosis of human choriocarcinoma cells. Biochem J 1 May 2001; 355 (3): 653–661. doi: https://doi.org/10.1042/bj3550653
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