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The ROS derived mitochondrial respirstion not from NADPH oxidase plays key role in Celastrol against angiotensin II-mediated HepG2 cell proliferation

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Abstract

Angiotensin II (AngII) is an important factor that promotes the proliferation of cancer cells, whereas celastrol exhibits a significant antitumor activity in various cancer models. Whether celastrol can effectively suppress AngII mediated cell proliferation remains unknown. In this study, we studied the effect of celastrol on AngII-induced HepG2 cell proliferation and evaluated its underlying mechanism. The results revealed that AngII was able to significantly promote HepG2 cell proliferation via up-regulating AngII type 1 (AT1) receptor expression, improving mitochondrial respiratory function, enhancing nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity, increasing the levels of reactive oxygen species (ROS) and pro-inflammatory cytokines. The excess ROS from mitochondrial dysfunction is able to cause the apoptosis of tumor cells via activating caspase3 signal pathway. In addition, the reaction between NO and ROS results in the formation of peroxynitrite (ONOO−), and then promoting cell damage. celastrol dramatically enhanced ROS generation, thereby causing cell apoptosis through inhibiting mitochodrial respiratory function and boosting the expression levels of AngII type 2 (AT2) receptor without influencing NADPH oxidase activity. PD123319 as a special inhibitor of AT2R was able to effectively decreased the levels of inflammatory cytokines and endothelial nitric oxide synthase (eNOS) activity, but only partially attenuate the effect of celastrol on AnII mediated HepG2 cell proliferation. Thus, celastrol has the potential for use in liver cancer therapy. ROS derived from mitochondrial is an important factor for celastrol to suppress HepG2 cell proliferation.

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Abbreviations

AngII:

Angiotensin II

NADPH oxidase:

Nicotinamide adenine dinucleotide phosphate oxidase

AT1R:

AngII type 1 receptor

AT2R:

AngII type 2 receptor

ROS:

Reactive oxygen species

MTT:

3-(4,5-Dimethythiazol-2-yl)-2,5-diphenyl-tetrazolium bromide

eNOS:

Endothelial nitric oxide synthase

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

EDTA:

Trypsin–ethylene diamine tetraacetic acid

VCAM:

Vascular cell adhesion molecule

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Acknowledgments

The authors thank Professor Lin at Xi’an Jiaotong University (Xi’an, Shanxi, China) for providing the HepG2 cell line. The authors would like to thank all members of the Department of Pharmacology, Binzhou Medical University for helpful discussions and technical assistance. The present study was supported by the Project of Shandong Province Higher Educational Science and Technology Program (No. J10LF89), the Project of Shandong Province Medicine & Health Science and Technology Program (2011HZ003), the Natural Science Foundation of Shandong Province (No. ZR2012HM076), and the Foundation of Taishan Scholar (tshw20110515).

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    Authors and Affiliations

    1. Key Laboratory of Traditional Chinese Medicine Prescription Effect and Clinical Evaluation of State Administration of Traditional Chinese Medicine, School of Pharmaceutical Sciences, Binzhou Medical University, Yantai, 264003, Shandong, China

      Xin Liu, Miao Li, Chun-feng Si, Yong-peng He, Min Wang, Ying Yang, Qing-yin Zheng & Chao-yun Wang

    2. Department of Pediatrics, Binzhou Medical University Hospital, Binzhou, 256600, Shandong, China

      Rui-wei Gao

    3. Department of Otolaryngology-Head & Neck Surgery, Case Western Reserve University, Cleveland, OH, 44106, USA

      Qing-yin Zheng

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    1. Xin Liu
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    Correspondence to Chao-yun Wang.

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    The authors declare no competing financial interests.

    Author contributions

    Participated in research designC.Y. Wang, Q.Y. Zheng. Conducted experiments: C.Y. Wang, X. Liu, R.W. Gao, M. Li, C.F. Si, Y.P. He, M. Wang, Y. Wu, Y.P. Zhang, Y. Yang. Performed data analysis: C.Y. Wang, X. Liu, R.W. Gao, J.R. Zhang. Wrote or contributed to the writing of the manuscript: C.Y. Wang, X. Liu, R.W. Gao.

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    Xin Liu and Rui-wei Gao have contributed equally to this work.

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    Liu, X., Gao, Rw., Li, M. et al. The ROS derived mitochondrial respirstion not from NADPH oxidase plays key role in Celastrol against angiotensin II-mediated HepG2 cell proliferation. Apoptosis 21, 1315–1326 (2016). https://doi.org/10.1007/s10495-016-1294-6

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