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Enhancement of hyperthermia-induced apoptosis by 5Z-7-oxozeaenol, a TAK1 inhibitor, in A549 cells

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Cell Stress and Chaperones Aims and scope

Abstract

KRAS mutant lung cancers have long been considered as untreatable with drugs. Transforming growth factor-β-activated kinase 1 (TAK1) appears to play an anti-apoptotic role in response to multiple stresses and has been reported to be a responsive kinase that regulates cell survival in KRAS-dependent cells. In this study, in order to find a useful approach to treat KRAS mutant lung cancer, we focused on the combined effects of 5Z-7-oxozeaenol, a TAK1 inhibitor, with hyperthermia (HT) in KRAS mutant lung cancer cell line A549. Annexin V-FITC/PI assay, cell cycle analysis, and colony formation assay revealed a significant enhancement in apoptosis induced by HT treatment, when the cells were pre-incubated with 5Z-7-oxozeaenol in a dose-dependent manner. The enhanced apoptosis by 5Z-7-oxozeaenol was accompanied by a significant increase in reactive oxygen species (ROS) generation and loss of mitochondrial membrane potential (MMP). In addition, western blot showed that 5Z-7-oxozeaenol enhanced HT-induced expressions of cleaved caspase-3, cleaved caspase-8, and HSP70 and decreased HT-induced expressions of Bcl-2, p-p38, p-JNK, and LC3. Moreover, 5Z-7-oxozeaenol pre-treatment resulted in a marked elevation of intracellular calcium level which might be associated with endoplasmic reticulum (ER) stress-related pathway. Taken together, our data provides further insights of the mechanism of action of 5Z-7-oxozeaenol and HT treatment, and their potential application as a novel approache to treat patients with KRAS mutant lung cancer.

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Abbreviations

AIF:

Apoptosis-inducing factor

DMSO:

Dimethyl sulfoxide

DiOC6(3):

3,3′-dihexyloxacarbocyanine iodide

FITC:

Fluorescein isothiocyanate

HE:

Hydroethidine

HPF:

Hydroxyphenyl fluorescein

HT:

Hyperthermia

IP3R:

Inositol 1,4,5-trisphosphate receptor

MAP3K:

Mitogen-activated protein kinase kinase kinase

MMP:

Mitochondrial membrane potential

MKKs:

Mitogen-activated protein kinase kinases

MPTP:

Mitochondria permeability transition pore

NSCLC:

Non-small cell lung cancer

PI:

Propidium iodide

ROS:

Reactive oxygen species

SCLC:

Small cell lung cancer

TAK1:

Transforming growth factor-β-activated kinase 1

References

  • Adams JM, Cory S (2001) Life-or-death decisions by the Bcl-2 protein family. Trends Biochem Sci 26:61–66

    Article  CAS  PubMed  Google Scholar 

  • Beere HM, Wolf BB, Cain K et al (2000) Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome. Nat Cell Biol 2:469–475

    Article  CAS  PubMed  Google Scholar 

  • Brookes PS, Yoon Y, Robotham JL, Anders MW, Sheu SS (2004) Calcium, ATP, and ROS: a mitochondrial love-hate triangle. Am J Physiol Cell Physiol 287:C817–833

    Article  CAS  PubMed  Google Scholar 

  • Chen F, Wang CC, Kim E, Harrison LE (2008) Hyperthermia in combination with oxidative stress induces autophagic cell death in HT-29 colon cancer cells. Cell Biol Int 32:715–723

    Article  CAS  PubMed  Google Scholar 

  • Chien CY, Chien CT, Wang SS (2014) Progressive thermopreconditioning attenuates rat cardiac ischemia/reperfusion injury by mitochondria-mediated antioxidant and antiapoptotic mechanisms. J Thorac Cardiovasc Surg 148:705–713

    Article  CAS  PubMed  Google Scholar 

  • Choi AY, Choi JH, Lee JY, Yoon KS, Choe W, Ha J et al (2010) Apigenin protects HT22 murine hippocampal neuronal cells against endoplasmic reticulum stress-induced apoptosis. Neurochem Int 57:143–152

    Article  CAS  PubMed  Google Scholar 

  • Cordero JB, Macagno JP, Stefanatos RK, Strathdee KE, Cagan RL, Vidal M (2010) Oncogenic Ras diverts a host TNF tumor suppressor activity into tumor promoter. Dev Cell 18:999–1011

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Daniel S, Arvelo MB, Patel VI, Longo CR, Shrikhande G, Shukri T et al (2004) A20 protects endothelial cells from TNF-, Fas-, and NK-mediated cell death by inhibiting caspase 8 activation. Blood 104:2376–2384

    Article  CAS  PubMed  Google Scholar 

  • Downward J (2003) Targeting RAS signalling pathways in cancer therapy. Nat Rev Cancer 3:11–22

    Article  CAS  PubMed  Google Scholar 

  • Engelhardt R (1987) Rational for clinical application of hyperthermia and drugs. Rev Med Chir Soc Med Nat Iasi 91:347–352

    CAS  PubMed  Google Scholar 

  • Fan Y, Cheng J, Vasudevan SA, Patel RH, Liang L, Xu X, Zhao Y, Jia W, Lu F, Zhang H, Nuchtern JG, Kim ES, Yang J (2013) TAK1 inhibitor 5Z-7-oxozeaenol sensitizes neuroblastoma to chemotherapy. Apoptosis 18:1224–1234

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Giorgi C, Baldassari F, Bononi A, Bonora M, De Marchi E, Marchi S, Missiroli S, Patergnani S, Rimessi A, Suski JM, Wieckowski MR, Pinton P (2012) Mitochondrial Ca(2+) and apoptosis. Cell Calcium 52:36–43

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gu ZT, Li L, Wu F, Zhao P, Yang H, Liu YS, Geng Y, Zhao M, Su L (2015) Heat stress induced apoptosis is triggered by transcription-independent p53, Ca(2+) dyshomeostasis and the subsequent Bax mitochondrial translocation. Sci Rep 5:11497

    Article  CAS  PubMed Central  Google Scholar 

  • Hayden MS, Ghosh S (2008) Shared principles in NF-kB signaling. Cell 132:344–362

    Article  CAS  PubMed  Google Scholar 

  • Hou CH, Lin FL, Hou SM, Liu JF (2014) Hyperthermia induces apoptosis through endoplasmic reticulum and reactive oxygen species in human osteosarcoma cells. Int J Mol Sci 15:17380–17395

    Article  PubMed  PubMed Central  Google Scholar 

  • Kim BJ, Ryu SW, Song BJ (2006) JNK- and p38 kinase-mediated phosphorylation of Bax leads to its activation and mitochondrial translocation and to apoptosis of human hepatoma HepG2 cells. J Biol Chem 281:21256–21265

    Article  CAS  PubMed  Google Scholar 

  • Ku BM, Jho EH, Bae YH, Sun JM, Ahn JS, Park K, Ahn MJ (2015) BYL719, a selective inhibitor of phosphoinositide 3-kinase α, enhances the effect of selumetinib (AZD6244, ARRY-142886) in KRAS-mutant non-small cell lung cancer. Invest New Drugs 33:12–21

    Article  CAS  PubMed  Google Scholar 

  • Lee CH, Jeon YT, Kim SH et al (2007) NF-kappaB as a potential molecular target for cancer therapy. BioFactors 29:19–35

    Article  CAS  PubMed  Google Scholar 

  • Li P, Furusawa Y, Wei ZL, Sakurai H, Tabuchi Y, Zhao QL, Saiki I, Kondo T (2013) TAK1 promotes cell survival by TNFAIP3 and IL-8 dependent and NF-kB independent pathway in HeLa cells exposed to heat stress. Int J Hyperthermia 29:688–695

    Article  PubMed  Google Scholar 

  • Li P, Zhao QL, Wu LH, Jawaid P, Jiao YF, Kadowaki M, Kondo T (2014) Isofraxidin, a potent reactive oxygen species (ROS) scavenger, protects human leukemia cells from radiation-induced apoptosis via ROS/mitochondria pathway in p53-independent manner. Apoptosis 19:1043–1053

    Article  CAS  PubMed  Google Scholar 

  • McAnulty SR, McAnulty L, Pascoe DD, Gropper SS, Keith RE, Morrow JD, Gladden LB (2005) Hyperthermia increases exercise-induced oxidative stress. Int J Sports Med 26:188–192

    Article  CAS  PubMed  Google Scholar 

  • McCullough KD, Martindale JL, Klotz LO, Aw TY, Holbrook NJ (2001) Gadd153 sensitizes cells to endoplasmic reticulum stress by down-regulating Bcl2 and perturbing the cellular redox state. Mol Cell Biol 21:1249–1259

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mihaly SR, Ninomiya-Tsuji J, Morioka S (2014) TAK1 control of cell death. Cell Death Differ 21:1667–1676

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Miyazaki Y, Kaikita K, Endo M, Horio E, Miura M, Tsujita K et al (2011) C/EBP homologous protein deficiency attenuates myocardial reperfusion injury by inhibiting myocardial apoptosis and inflammation. Arterioscler Thromb Vasc Biol 31:1124–1132

    Article  CAS  PubMed  Google Scholar 

  • Neubert M, Ridder DA, Bargiotas P, Akira S, Schwaninger M (2011) Acute inhibition of TAK1 protects against neuronal death in cerebral ischemia. Cell Death Differ 18:1521–1530

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ninomiya-Tsuji J, Kajino T, Ono K, Ohtomo T, Matsumoto M, Shiina M, Mihara M, Tsuchiya M, Matsumoto K (2003) A resorcylic acid lactone, 5Z-7-oxozeaenol, prevents inflammation by inhibiting the catalytic activity of TAK1 MAPK kinase kinase. J Biol Chem 278:18485–18490

    Article  CAS  PubMed  Google Scholar 

  • Omori E, Morioka S, Matsumoto K, Ninomiya-Tsuji J (2008) TAK1 regulates reactive oxygen species and cell death in keratinocytes, which is essential for skin integrity. J Biol Chem 283:26161–26168

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Orrenius S, Zhivotovsky B, Nicotera P (2003) Regulation of cell death: the calcium-apoptosis link. Nat Rev Mol Cell Biol 4:552–565

    Article  CAS  PubMed  Google Scholar 

  • Ott M, Gogvadze V, Orrenius S, Zhivotovsky B (2007) Mitochondria, oxidative stress and cell death. Apoptosis 12:913–922

    Article  CAS  PubMed  Google Scholar 

  • Oyadomari S, Mori M (2004) Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ 11:381–389

    Article  CAS  PubMed  Google Scholar 

  • Sakurai H (2012) Targeting of TAK1 in inflammatory disorders and cancer. Trends Pharmacol Sci 33:522–530

    Article  CAS  PubMed  Google Scholar 

  • Schober A (2008) Chemokines in vascular dysfunction and remodeling. Arterioscler Thromb Vasc Biol 28:1950–1959

    Article  CAS  PubMed  Google Scholar 

  • Siegel R, Ma J, Zou Z, Jemal A (2014) Cancer statistics. CA Cancer J Clin 64:9–29

    Article  PubMed  Google Scholar 

  • Singh A, Sweeney MF, Yu M, Burger A, Greninger P, Benes C, Haber DA, Settleman J (2012) TAK1 inhibition promotes apoptosis in KRAS-dependent colon cancers. Cell 148:639–650

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Song Z, Zhu X, Jin R, Wang C, Yan J, Zheng Q, Nanda A, Granger DN, Li G (2014) Roles of the kinase TAK1 in CD40-mediated effects on vascular oxidative stress and neointima formation after vascular injury. PLoS One 9:e101671

    Article  PubMed  PubMed Central  Google Scholar 

  • Speidel D (2010) Transcription-independent p53 apoptosis: an alternative route to death. Trends Cell Biol 20:14–24

    Article  CAS  PubMed  Google Scholar 

  • Swanlund JM, Kregel KC, Oberley TD (2008) Autophagy following heat stress: the role of aging and protein nitration. Autophagy 4:936–939

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tripathi P, Hildeman D (2004) Sensitization of T cells to apoptosis—a role for ROS? Apoptosis 9:515–523

    Article  CAS  PubMed  Google Scholar 

  • Turrens JF (2003) Mitochondrial formation of reactive oxygen species. J Physiol 555:335–344

    Article  Google Scholar 

  • Wold LE, Ceylan-Isik AF, Fang CX, Yang X, Li SY, Sreejayan N, Privratsky JR, Ren J (2006) Metallothionein alleviates cardiac dysfunction in streptozotocin-induced diabetes: role of Ca2+ cycling proteins, NADPH oxidase, poly(ADP-Ribose) polymerase and myosin heavy chain isozyme. Free Radic Biol Med 40:1419–1429

    Article  CAS  PubMed  Google Scholar 

  • Yamaguchi K, Shirakabe K, Shibuya H, Irie K, Oishi I, Ueno N et al (1995) Identification of a member of the MAPKKK family as a potential mediator of TGF-β signal transduction. Science 270:2008–2011

    Article  CAS  PubMed  Google Scholar 

  • Yin XM (2000) Signal transduction mediated by Bid, a pro-death Bcl-2 family proteins, connects the death receptor and mitochondria apoptosis pathways. Cell Res 10:161–167

    Article  CAS  PubMed  Google Scholar 

  • Yu T, Robotham JL, Yoon Y (2006) Increased production of reactive oxygen species in hyperglycemic conditions requires dynamic change of mitochondrial morphology. Proc Natl Acad Sci U S A 103:2653–2658

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang Y, Calderwood SK (2011) Autophagy, protein aggregation and hyperthermia: a mini-review. Int J Hyperthermia 27:409–414

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhao QL, Fujiwara Y, Kondo T (2006) Mechanism of cell death induction by nitroxide and hyperthermia. Free Radic Biol Med 40:1131–1143

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported in part by Grant-in-Aid for Challenging Exploratory Research (No. 26670551), Japan society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Radiological Sciences, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Sugitani 2630, Toyama, 930-0194, Japan

    Peng Li, Qing-Li Zhao, Paras Jawaid, Mati Ur Rehman & Takashi Kondo

  2. Department of Cancer Cell Biology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Sugitani 2630, Toyama, 930-0194, Japan

    Hiroaki Sakurai

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  1. Peng Li
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  2. Qing-Li Zhao
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  3. Paras Jawaid
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  6. Takashi Kondo
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Correspondence to Qing-Li Zhao.

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Li, P., Zhao, QL., Jawaid, P. et al. Enhancement of hyperthermia-induced apoptosis by 5Z-7-oxozeaenol, a TAK1 inhibitor, in A549 cells. Cell Stress and Chaperones 21, 873–881 (2016). https://doi.org/10.1007/s12192-016-0712-6

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  • DOI: https://doi.org/10.1007/s12192-016-0712-6

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