Full Text:   <375>

Summary:  <117>

CLC number: R73-3

On-line Access: 2019-04-01

Received: 2018-06-07

Revision Accepted: 2018-11-15

Crosschecked: 2019-03-02

Cited: 0

Clicked: 630

Citations:  Bibtex RefMan EndNote GB/T7714


Hui Pan


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Journal of Zhejiang University SCIENCE B 2019 Vol.20 No.4 P.310-321


Mitochondrial superoxide anions induced by exogenous oxidative stress determine tumor cell fate: an individual cell-based study

Author(s):  Hui Pan, Bao-hui Wang, Zhou-bin Li, Xing-guo Gong, Yong Qin, Yan Jiang, Wei-li Han

Affiliation(s):  The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China; more

Corresponding email(s):   phwbhpxc@zju.edu.cn, wang.baohui888@163.com, zjhzhwldoc@zju.edu.cn

Key Words:  Individual cell, Superoxide anion, Reactive oxygen species (ROS) dynamics, Intrinsic apoptotic pathway, Ca2+ signaling

Hui Pan, Bao-hui Wang, Zhou-bin Li, Xing-guo Gong, Yong Qin, Yan Jiang, Wei-li Han. Mitochondrial superoxide anions induced by exogenous oxidative stress determine tumor cell fate: an individual cell-based study[J]. Journal of Zhejiang University Science B, 2019, 20(4): 310-321.

@article{title="Mitochondrial superoxide anions induced by exogenous oxidative stress determine tumor cell fate: an individual cell-based study",
author="Hui Pan, Bao-hui Wang, Zhou-bin Li, Xing-guo Gong, Yong Qin, Yan Jiang, Wei-li Han",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Mitochondrial superoxide anions induced by exogenous oxidative stress determine tumor cell fate: an individual cell-based study
%A Hui Pan
%A Bao-hui Wang
%A Zhou-bin Li
%A Xing-guo Gong
%A Yong Qin
%A Yan Jiang
%A Wei-li Han
%J Journal of Zhejiang University SCIENCE B
%V 20
%N 4
%P 310-321
%@ 1673-1581
%D 2019
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1800319

T1 - Mitochondrial superoxide anions induced by exogenous oxidative stress determine tumor cell fate: an individual cell-based study
A1 - Hui Pan
A1 - Bao-hui Wang
A1 - Zhou-bin Li
A1 - Xing-guo Gong
A1 - Yong Qin
A1 - Yan Jiang
A1 - Wei-li Han
J0 - Journal of Zhejiang University Science B
VL - 20
IS - 4
SP - 310
EP - 321
%@ 1673-1581
Y1 - 2019
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1800319

Objective: Reactive oxygen species (ROS) are involved in a variety of biological phenomena and serve both deleterious and beneficial roles. ROS quantification and assessment of reaction networks are desirable but difficult because of their short half-life and high reactivity. Here, we describe a pro-oxidative model in a single human lung carcinoma SPC-A-1 cell that was created by application of extracellular H2O2 stimuli. Methods: Modified microfluidics and imaging techniques were used to determine O2•− levels and construct an O2•− reaction network. To elucidate the consequences of increased O2•− input, the mitochondria were given a central role in the oxidative stress mode, by manipulating mitochondria-interrelated cytosolic Ca2+ levels, mitochondrial Ca2+ uptake, auto-amplification of intracellular ROS and the intrinsic apoptotic pathway. Results and conclusions: Results from a modified microchip demonstrated that 1 mmol/L H2O2 induced a rapid increase in cellular O2•− levels (>27 vs. >406 amol in 20 min), leading to increased cellular oxidizing power (evaluated by ROS levels) and decreased reducing power (evaluated by glutathione (GSH) levels). In addition, we examined the dynamics of cytosolic Ca2+ and mitochondrial Ca2+ by confocal laser scanning microscopy and confirmed that Ca2+ stores in the endoplasmic reticulum were the primary source of H2O2-induced cytosolic Ca2+ bursts. It is clear that mitochondria have pivotal roles in determining how exogenous oxidative stress affects cell fate. The stress response involves the transfer of Ca2+ signals between organelles, ROS auto-amplification, mitochondrial dysfunction, and a caspase-dependent apoptotic pathway.


结论:研究结果表明1 mmol/L H2O2引起细胞O2•−水平的快速增加,从而导致细胞氧化能力增加和还原能力降低.此外,研究还证实了内质网中储存的Ca2+是H2O2诱导的线粒体Ca2+爆发的主要来源.外源氧化压力反应涉及细胞器间Ca2+信号的传递、ROS自身扩增、线粒体功能紊乱和半胱天冬酶依赖性凋亡途径.线粒体在外源性氧化应激影响细胞命运方面发挥着关键作用.


Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Akopova OV, 2008. The role of mitochondrial permeability transition pore in transmembrane Ca2+-exchange in mitochondria. Ukr Biokhim Zh (1999), 80(3):40-47 (in Ukrainian).

[2]Blokhina O, Fagerstedt KV, 2010. Oxidative metabolism, ROS and NO under oxygen deprivation. Plant Physiol Biochem, 48(5):359-373.

[3]Bogeski I, Kappl R, Kummerow C, et al., 2011. Redox regulation of calcium ion channels: chemical and physiological aspects. Cell Calcium, 50(5):407-423.

[4]Camello-Almaraz C, Gomez-Pinilla PJ, Pozo MJ, et al., 2006. Mitochondrial reactive oxygen species and Ca2+ signaling. Am J Physiol Cell Physiol, 291(5):C1082-C1088.

[5]de Marchi E, Bonora M, Giorgi C, et al., 2014. The mitochondrial permeability transition pore is a dispensable element for mitochondrial calcium efflux. Cell Calcium, 56(1):1-13.

[6]Feng W, Liu GH, Allen PD, et al., 2000. Transmembrane redox sensor of ryanodine receptor complex. J Biol Chem, 275(46):35902-35907.

[7]Gao J, Yin XF, Fang ZL, 2004. Integration of single cell injection, cell lysis, separation and detection of intracellular constituents on a microfluidic chip. Lab Chip, 4(1):47-52.

[8]Gao N, Li L, Shi ZK, et al., 2007. High-throughput determination of glutathione and reactive oxygen species in single cells based on fluorescence images in a microchannel. Electrophoresis, 28(21):3966-3975.

[9]Giorgi C, Missiroli S, Patergnani S, et al., 2015. Mitochondria-associated membranes: composition, molecular mechanisms, and physiopathological implications. Antioxid Redox Signal, 22(12):995-1019.

[10]Hileman EO, Liu JS, Albitar M, et al., 2004. Intrinsic oxidative stress in cancer cells: a biochemical basis for therapeutic selectivity. Cancer Chemother Pharmacol, 53(3):209-219.

[11]Köhler AC, Sag CM, Maier LS, 2014. Reactive oxygen species and excitation-contraction coupling in the context of cardiac pathology. J Mol Cell Cardiol, 73:92-102.

[12]Labuschagne CF, Brenkman AB, 2013. Current methods in quantifying ROS and oxidative damage in Caenorhabditis elegans and other model organism of aging. Ageing Res Rev, 12(4):918-930.

[13]Liochev SI, 2013. Reactive oxygen species and the free radical theory of aging. Free Radic Biol Med, 60:1-4.

[14]Lyublinskaya OG, Zenin VV, Shatrova AN, et al., 2014. Intracellular oxidation of hydroethidine: compartmentalization and cytotoxicity of oxidation products. Free Radic Biol Med, 75:60-68.

[15]Maltepe E, Saugstad OD, 2009. Oxygen in health and disease: regulation of oxygen homeostasis-clinical implications. Pediatr Res, 65(3):261-268.

[16]Martindale JL, Holbrook NJ, 2002. Cellular response to oxidative stress: signaling for suicide and survival. J Cell Physiol, 192(1):1-15.

[17]Moreau B, Nelson C, Parekh AB, 2006. Biphasic regulation of mitochondrial Ca2+ uptake by cytosolic Ca2+ concentration. Curr Biol, 16(16):1672-1677.

[18]Mota SI, Costa RO, Ferreira IL, et al., 2015. Oxidative stress involving changes in Nrf2 and ER stress in early stages of Alzheimer’s disease. Biochim Biophys Acta, 1852(7):1428-1441.

[19]Orrenius S, Gogvadze V, Zhivotovsky B, 2015. Calcium and mitochondria in the regulation of cell death. Biochem Biophys Res Commun, 460(1):72-81.

[20]Pan H, Wang BH, Lv W, et al., 2015. Esculetin induces apoptosis in human gastric cancer cells through a cyclophilin D-mediated mitochondrial permeability transition pore associated with ROS. Chemico-Biological Interactions, 242:51-60.

[21]Pelicano H, Carney D, Huang P, 2004. ROS stress in cancer cells and therapeutic implications. Drug Resist Updat, 7(2):97-110.

[22]Qin Y, Chen FD, Zhou L, et al., 2009. Proliferative and anti-proliferative effects of thymosin α1 on cells are associated with manipulation of cellular ROS levels. Chem Biol Interact, 180(3):383-388.

[23]Qin Y, Pan X, Tang TT, et al., 2011. Anti-proliferative effects of the novel squamosamide derivative (FLZ) on HepG2 human hepatoma cells by regulating the cell cycle-related proteins are associated with decreased Ca2+/ROS levels. Chem Biol Interact, 193(3):246-253.

[24]Rhee SG, 2006. Cell signaling: H2O2, a necessary evil for cell signaling. Science, 312(5782):1882-1883.

[25]Singh DK, Kumar D, Siddiqui Z, et al., 2005. The strength of receptor signaling is centrally controlled through a cooperative loop between Ca2+ and an oxidant signal. Cell, 121(2):281-293.

[26]Solier S, Pommier Y, 2011. MDC1 cleavage by caspase-3: a novel mechanism for inactivating the DNA damage response during apoptosis. Cancer Res, 71(3):906-913.

[27]Spät A, Pitter JG, 2004. The effect of cytoplasmic Ca2+ signal on the redox state of mitochondrial pyridine nucleotides. Mol Cell Endocrinol, 215(1-2):115-118.

[28]Sun Y, Yin XF, 2006. Novel multi-depth microfluidic chip for single cell analysis. J Chromatogr A, 1117(2):228-233.

[29]Sun Y, Yin XF, Ling YY, et al., 2005. Determination of reactive oxygen species in single human erythrocytes using microfluidic chip electrophoresis. Anal Bioanal Chem, 382(7):1472-1476.

[30]Tang HY, Qin Y, Li JY, et al., 2011. The scavenging of superoxide radicals promotes apoptosis induced by a novel cell-permeable fusion protein, sTRAIL:FeSOD, in tumor necrosis factor-related apoptosis-inducing ligand-resistant leukemia cells. BMC Biol, 9:18.

[31]Thannickal VJ, Fanburg BL, 2000. Reactive oxygen species in cell signaling. Am J Physiol Lung Cell Mol Physiol, 279(6):L1005-L1028.

[32]Tonks NK, 2005. Redox redux: revisiting PTPs and the control of cell signaling. Cell, 121(5):667-670.

[33]von Montfort C, Matias N, Fernandez A, et al., 2012. Mitochondrial GSH determines the toxic or therapeutic potential of superoxide scavenging in steatohepatitis. J Hepatol, 57(4):852-859.

[34]Voronina S, Sukhomlin T, Johnson PR, et al., 2002. Correlation of NADH and Ca2+ signals in mouse pancreatic acinar cells. J Physiol, 539:41-52.

[35]Yu SY, Jang Y, Paik D, et al., 2015. Nmdmc overexpression extends Drosophila lifespan and reduces levels of mitochondrial reactive oxygen species. Biochem Biophys Res Commun, 465(4):845-850.

[36]Yuana Y, Sturk A, Nieuwland R, 2013. Extracellular vesicles in physiological and pathological conditions. Blood Rev, 27(1):31-39.

[37]Zhao HT, Kalivendi S, Zhang H, et al., 2003. Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: potential implications in intracellular fluorescence detection of superoxide. Free Radic Biol Med, 34(11):1359-1368.

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