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High dose of extracellular ATP switched autophagy to apoptosis in anchorage-dependent and anchorage-independent hepatoma cells

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Abstract

Extracellular adenosine triphosphate (eATP) transduces purinergic signal and plays an important regulatory role in many biological processes, including tumor cell growth and cell death. A large amount of eATP exists in the fast-growing tumor center and inflammatory tumor microenvironment. Tumor cells could acquire anoikis resistance and anchorage independence in tumor microenvironment and further cause metastatic lesion. Whether such a high amount of eATP has any effect on the anchored and non-anchored tumor cells in tumor microenvironment has not been elucidated and is investigated in this study. Our data showed that autophagy helped hepatoma cells to maintain survival under the treatment of no more than 1 mM of eATP. Only when eATP concentration reached a relatively high level (2.5 mM), cell organelle could not be further maintained by autophagy, and apoptosis and cell death occurred. In hepatoma cells under treatment of 2.5 mM of eATP, an AMP-activated protein kinase (AMPK) pathway was dramatically activated while mTOR signaling pathway was suppressed in coordination with apoptosis. Further investigation showed that the AMPK/mTOR axis played a key role in tipping the balance between autophagy-mediated cell survival and apoptosis-induced cell death under the treatment of eATP. This work provides evidence to explain how hepatoma cells escape from eATP-induced cytotoxicity as well as offers an important clue to consider effective manipulation of cancer.

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Abbreviations

ATP:

Adenosine triphosphate

eATP:

Extracellular adenosine triphosphate

PCR:

Polymerase chain reaction

RT-PCR:

Reverse transcriptase polymerase chain reaction

3-MA:

3-Methyladenine

LC3:

Microtubule-associated protein light chain 3

CCK-8:

Cell Counting Kit-8

Min:

Minutes

H:

Hours

mTOR:

Mammalian target of rapamycin

S6K1:

Ribosomal S6 kinase 1

PBS:

Phosphate-buffered saline

FBS:

Fetal bovine serum

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Acknowledgments

This study was supported by the National Nature Science Foundation of China (No. 81172352, No. 30700357), Science and Technology Development Project of Shandong Province (2011GGE27020), and Project for Outstanding Young Scientist in Jinan City (No.20080211 ).

Conflict of interest

The authors declare no conflict of interests.

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Authors and Affiliations

  1. Department of Immunology, Shandong Provincial Key Laboratory of Infection & Immunology, School of Medicine, Shandong University, 44 West Wenhua Road, Jinan, 250012, China

    Qing Wei, Ying Zhang, Lei Sun, Wanwan Huai, Zhengkun Wan & Lihui Han

  2. Shandong University Qilu Hospital, Jinan, 250012, China

    Xiaoqing Jia

  3. Shengli Hospital (affiliated with Shandong University), Jinan, 250021, China

    Chengcheng Yu

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  1. Qing Wei
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Correspondence to Lihui Han.

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Wei, Q., Zhang, Y., Sun, L. et al. High dose of extracellular ATP switched autophagy to apoptosis in anchorage-dependent and anchorage-independent hepatoma cells. Purinergic Signalling 9, 585–598 (2013). https://doi.org/10.1007/s11302-013-9369-0

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  • DOI: https://doi.org/10.1007/s11302-013-9369-0

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