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Clitocine targets Mcl-1 to induce drug-resistant human cancer cell apoptosis in vitro and tumor growth inhibition in vivo

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Abstract

Drug resistance is a major reason for therapy failure in cancer. Clitocine is a natural amino nucleoside isolated from mushroom and has been shown to inhibit cancer cell proliferation in vitro. In this study, we observed that clitocine can effectively induce drug-resistant human cancer cell apoptosis in vitro and inhibit tumor xenograft growth in vivo. Clitocine treatment inhibited drug-resistant human cancer cell growth in vitro in a dose- and time-dependent manner. Biochemical analysis revealed that clitocine-induced tumor growth inhibition is associated with activation of caspases 3, 8 and 9, PARP cleavage, cytochrome c release and Bax, Bak activation, suggesting that clitocine inhibits drug-resistant cancer cell growth through induction of apoptosis. Analysis of apoptosis regulatory genes indicated that Mcl-1 level was dramatically decreased after clitocine treatment. Over-expression of Mcl-1 reversed the activation of Bax and attenuated clitocine-induced apoptosis, suggesting that clitocine-induced apoptosis was at least partially by inducing Mcl-1 degradation to release Bax and Bak. Consistent with induction of apoptosis in vitro, clitocine significantly suppressed the drug-resistant hepatocellular carcinoma xenograft growth in vivo by inducing apoptosis as well as inhibiting cell proliferation. Taken together, our data demonstrated that clitocine is a potent Mcl-1 inhibitor that can effectively induce apoptosis to suppress drug-resistant human cancer cell growth both in vitro and in vivo, and thus holds great promise for further development as potentially a novel therapeutic agent to overcome drug resistance in cancer therapy.

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Abbreviations

HCC:

Human hepatocellular carcinoma

BCL-2:

B-cell lymphoma-2

Mcl-1:

Myeloid cell leukaemia-1

Bak:

Bcl-2 homologous antagonist-killer

Bax:

Bcl-2-associated X protein

ΔΨm :

Mitochondrial transmembrane potential

PARP:

Poly (ADP) ribose polymerase

P-gp:

P-glycoprotein

NF-κB:

Nuclear factor-kappa B

DOX:

Doxorubicin

IHC:

Immunohistochemistry

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining

IC50 :

Concentration causing a 50 % growth inhibition

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Acknowledgments

This work was supported in part by the Nature Science Foundation of Zhejiang Province (Grant No.: LY12C02001 to FL) and Science and Technology Department of Zhejiang Province (Grant No.: 2013C24012 to FL), and Siyuan Foundation (to FL and KPF). We thank Dr. Shihua Wu for assistance in purification of Clitocine.

Conflicts of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Research Centre of Siyuan Natural Pharmacy and Biotoxicology, College of Life Sciences, Zhejiang University, Zi**ggang Campus, Hangzhou, 310058, People’s Republic of China

    Jian-guo Sun, Hua Li, **a Li, Xueli Zeng, ** Wu & Fei-yan Liu

  2. Joint Centre of Zhejiang University and The Chinese University of Hong Kong on Natural Products and Toxicology Research, Zhejiang University, Zi**ggang Campus, Hangzhou, People’s Republic of China

    Jian-guo Sun, ** Wu, Kwok-Pui Fung & Fei-yan Liu

  3. The State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Zi**ggang Campus, Hangzhou, People’s Republic of China

    ** Wu

  4. School of Biomedical Sciences (SBS), The Chinese University of Hong Kong, Shatin, Hong Kong SAR, People’s Republic of China

    Kwok-Pui Fung

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  1. Jian-guo Sun
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  2. Hua Li
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Correspondence to Fei-yan Liu.

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Sun, Jg., Li, H., Li, X. et al. Clitocine targets Mcl-1 to induce drug-resistant human cancer cell apoptosis in vitro and tumor growth inhibition in vivo. Apoptosis 19, 871–882 (2014). https://doi.org/10.1007/s10495-014-0969-0

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