Skip to main content
SpringerLink
Log in

Tunicamycin promotes apoptosis in leukemia cells through ROS generation and downregulation of survivin expression

  • Original Paper
  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

Tunicamycin (TN), one of the endoplasmic reticulum stress inducers, has been reported to inhibit tumor cell growth and exhibit anticarcinogenic activity. However, the mechanism by which TN initiates apoptosis remains poorly understood. In the present study, we investigated the effect of TN on the apoptotic pathway in U937 cells. We show that TN induces apoptosis in association with caspase-3 activation, generation of reactive oxygen species (ROS), and downregulation of survivin expression. P38 MAPK (mitogen-activated protein kinase) and the generation of ROS signaling pathway play crucial roles in TN-induced apoptosis in U937 cells. We hypothesized that TN-induced activation of p38 MAPK signaling pathway is responsible for cell death. To test this hypothesis, we selectively inhibited MAPK during treatment with TN. Our data demonstrated that inhibitor of p38 (SB), but not ERK (PD) or JNK (SP), partially maintained apoptosis during treatment with TN. Pre-treatment with NAC and GSH markedly prevented cell death, suggesting a role for ROS in this process. Ectopic expression of survivin in U937 cells attenuated TN-induced apoptosis by suppression of caspase-3 cleavage, mitochondrial membrane potential, and cytochrome c release in U937 cells. Taken together, our results show that TN modulates multiple components of the apoptotic response of human leukemia cells and raise the possibility of a novel therapeutic strategy for hematological malignancies.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

DAPI:

4′,6′-Diamidino-2-phenylindole

DCFDA:

2′7′-Dichlorodihydrofluorescein diacetate

NAC:

N-acetylcysteine

PBS:

Phosphate-buffered saline

PI:

Propidium iodide

ROS:

Reactive oxygen species

UPR:

Unfolded protein response

z-VAD-fmk:

Benzyl carbonyl-Val-Ala-Asp-fluoromethyl ketone

References

  1. Noda I, Fujieda S, Seki M, Tanaka N, Sunaga H, Ohtsubo T, Tsuzuki H, Fan GK, Saito H (1999) Inhibition of N-linked glycosylation by tunicamycin enhances sensitivity to cisplatin in human head-and-neck carcinoma cells. Int J Cancer 80:279–284

    Article  CAS  PubMed  Google Scholar 

  2. Dennis JW, Granovsky M, Warren CE (1999) Protein glycosylation in development and disease. BioEssays 21:412–421

    Article  CAS  PubMed  Google Scholar 

  3. Goss PE, Baker MA, Carver JP, Dennis JW (1995) Inhibitors of carbohydrate processing: a new class of anticancer agents. Clin Cancer Res 1:935–944

    CAS  PubMed  Google Scholar 

  4. Bentley J, Quinn DM, Pitman RS, Warr JR, Kellett GL (1997) The human KB multidrug-resistant cell line KB-C1 is hypersensitive to inhibitors of glycosylation. Cancer Lett 115:221–227

    Article  CAS  PubMed  Google Scholar 

  5. Carlberg M, Larsson O (1993) Role of N-linked glycosylation in cell-cycle progression and initiation of DNA synthesis in tumor-transformed human fibroblasts. Anticancer Res 13:167–171

    CAS  PubMed  Google Scholar 

  6. Martínez JA, Torres-Negrón I, Amigó LA, Banerjee DK (1999) Expression of Glc3Man9GlcNAc2-PP-Dol is a prerequisite for capillary endothelial cell proliferation. Cell Mol Biol (Noisy-le-grand) 45:137–152

    Google Scholar 

  7. Hiss D, Gabriels G, Jacobs P, Folb P (1996) Tunicamycin potentiates drug cytotoxicity and vincristine retention in multidrug resistant cell lines. Eur J Cancer 32:2164–2172

    Article  Google Scholar 

  8. Kramer R, Weber TK, Arceci R, Ramchurren N, Kastrinakis WV, Steele G Jr, Summerhayes IC (1995) Inhibition of N-linked glycosylation of P-glycoprotein by tunicamycin results in a reduced multidrug resistance phenotype. Br J Cancer 71:670–675

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. Lou LX, Geng B, Yu F, Zhang J, Pan CS, Chen L, Qi YF, Ke Y, Wang X, Tang CS (2006) Endoplasmic reticulum stress response is involved in the pathogenesis of stress induced gastric lesions in rats. Life Sci 79:1856–1864

    Article  CAS  PubMed  Google Scholar 

  10. Rao RV, Poksay KS, Castro-Obregon S, Schilling B, Row RH, del Rio G, Gibson BW, Ellerby HM, Bredesen DE (2004) Molecular components of a cell death pathway activated by endoplasmic reticulum stress. J Biol Chem 279:177–187

    Article  CAS  PubMed  Google Scholar 

  11. Reimertz C, Kögel D, Rami A, Chittenden T, Prehn JH (2003) Gene expression during ER stress-induced apoptosis in neurons: induction of the BH3-only protein Bbc3/PUMA and activation of the mitochondrial apoptosis pathway. J Cell Biol 162:587–597

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  12. Walker BK, Lei H, Krag SS (1998) A functional link between N-linked glycosylation and apoptosis in Chinese hamster ovary cells. Biochem Biophys Res Commun 250:264–270

    Article  CAS  PubMed  Google Scholar 

  13. Kleizen B, Braakman I (2004) Protein folding and quality control in the endoplasmic reticulum. Curr Opin Cell Biol 16:343–349

    Article  CAS  PubMed  Google Scholar 

  14. Aridor M, Balch WE (1999) Integration of endoplasmic reticulum signaling in health and disease. Nat Med 5:745–751

    Article  CAS  PubMed  Google Scholar 

  15. Xu C, Bailly-Maitre B, Reed JC (2005) Endoplasmic reticulum stress: cell life and death decisions. J Clin Invest 115:2656–2664

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  16. Kim I, Xu W, Reed JC (2008) Cell death and endoplasmic reticulum stress: disease relevance and herapeutic opportunities. Nat Rev Drug Discov 7:1013–1030

    Article  CAS  PubMed  Google Scholar 

  17. Rutkowski DT, Kaufman RJ (2004) A trip to the ER: coping with stress. Trends Cell Biol 14:20–28

    Article  CAS  PubMed  Google Scholar 

  18. Elbein AD (1987) Inhibitors of the biosynthesis and processing of N-linked oligosaccharide chains. Annu Rev Biochem 56:497–534

    Article  CAS  PubMed  Google Scholar 

  19. Natsume Y, Ito S, Satsu H, Shimizu M (2009) Protective effect of quercetin on ER stress caused by calcium dynamics dysregulation in intestinal epithelial cells. Toxicology 258:164–175

    Article  CAS  PubMed  Google Scholar 

  20. Xu C, Bailly-Maitre B, Reed JC (2005) Endoplasmic reticulum stress: cell life and death decisions. J Clin Invest 115:2656–2664

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  21. Hattori M, Sakamoto H, Satoh K, Yamamoto T (2001) DNA demethylase is expressed in ovarian cancers and the expression correlates with demethylation of CpG sites in the promoter region of c-erbB-2 and survivin genes. Cancer Lett 169:155–164

    Article  CAS  PubMed  Google Scholar 

  22. Li F, Altieri DC (1999) Transcriptional analysis of human survivin gene expression. Biochem J 344Pt 2:305–311

  23. Hoffman WH, Biade S, Zilfou JT, Chen J, Murphy M (2002) Transcriptional repression of the anti-apoptotic survivin gene by wild type p53. J Biol Chem 277:3247–3257

    Article  CAS  PubMed  Google Scholar 

  24. Mirza A, McGuirk M, Hockenberry TN, Wu Q, Ashar H, Black S, Wen SF, Wang L, Kirschmeier P, Bishop WR, Nielsen LL, Pickett CB, Liu S (2002) Human survivin is negatively regulated by wild-type p53 and participates in p53-dependent apoptotic pathway. Oncogene 21:2613–2622

    Article  CAS  PubMed  Google Scholar 

  25. Simoncini S, Sapet C, Camoin-Jau L, Bardin N, Harlé JR, Sampol J, Dignat-George F, Anfosso F (2005) Role of reactive oxygen species and p38 MAPK in the induction of the pro-adhesive endothelial state mediated by IgG from patients with anti-phospholipid syndrome. Int Immunol 17:489–500

    Article  CAS  PubMed  Google Scholar 

  26. Kim BM, Lee KH, Hong IS, Hong SH (2012) p38 mitogen-activated protein kinase is a key regulator of 5-phenylselenyl- and 5-methylselenyl-methyl-2′-deoxyuridine-induced apoptosis in human HL-60 cells. Biochem Biophys Res Commun 417:237–244

    Article  CAS  PubMed  Google Scholar 

  27. Kim MJ, Lee TH, Kim SH, Choi YJ, Heo J, Kim YH (2010) Triptolide inactivates Akt and induces caspase-dependent death in cervical cancer cells via the mitochondrial pathway. Int J Oncol 37:1177–1185

    CAS  PubMed  Google Scholar 

  28. Fleury C, Mignotte B, Vayssière JL (2002) Mitochondrial reactive oxygen species in cell death signaling. Biochimie 84:131–141

    Article  CAS  PubMed  Google Scholar 

  29. Wen J, You KR, Lee SY, Song CH, Kim DG (2002) Oxidative stress-mediated apoptosis. The anticancer effect of the sesquiterpene lactone parthenolide. J Biol Chem 277:38954–38964

    Article  CAS  PubMed  Google Scholar 

  30. Jin HO, Yoon SI, Seo SK, Lee HC, Woo SH, Yoo DH, Lee SJ, Choe TB, An S, Kwon TJ, Kim JI, Park MJ, Hong SI, Park IC, Rhee CH (2006) Synergistic induction of apoptosis by sulindac and arsenic trioxide in human lung cancer A549 cells via reactive oxygen species-dependent down-regulation of survivin. Biochem Pharmacol 72:1228–1236

    Article  CAS  PubMed  Google Scholar 

  31. Guan L, Han B, Li Z, Hua F, Huang F, Wei W, Yang Y, Xu C (2009) Sodium selenite induces apoptosis by ROS-mediated endoplasmic reticulum stress and mitochondrial dysfunction in human acute promyelocytic leukemia NB4 cells. Apoptosis 14:218–225

    Article  CAS  PubMed  Google Scholar 

  32. Ono Y, Shimazawa M, Ishisaka M, Oyagi A, Tsuruma K, Hara H (2012) Imipramine protects mouse hippocampus against tunicamycin-induced cell death. Eur J Pharmacol 696:83–88

    Article  CAS  PubMed  Google Scholar 

  33. Shore GC, Papa FR, Oakes SA (2011) Signaling cell death from the endoplasmic reticulum stress response. Curr Opin Cell Biol 23:143–149

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  34. Okada K, Minamino T, Tsukamoto Y, Liao Y, Tsukamoto O, Takashima S, Hirata A, Fujita M, Nagamachi Y, Nakatani T (2004) Prolonged endoplasmic reticulum stress in hypertrophic and failing heart after aortic constriction: possible contribution of endoplasmic reticulum stress to cardiac myocyte apoptosis. Circulation 110:705–712

    Article  PubMed  Google Scholar 

  35. Zhao H, Liao Y, Minamino T, Asano Y, Asakura M, Kim J, Asanuma H, Takashima S, Hori M, Kitakaze M (2008) Inhibition of cardiac remodeling by pravastatin is associated with amelioration of endoplasmic reticulum stress. Hypertens Res 31:1977–1987

    Article  CAS  PubMed  Google Scholar 

  36. Marciniak SJ, Yun CY, Oyadomari S, Novoa I, Zhang Y, Jungreis R, Nagata K, Harding HP, Ron D (2004) CHOP induces death by promoting protein synthesis and oxidation in the stressed endoplasmic reticulum. Genes Dev 18:3066–3077

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  37. Li Y, Xie M, Yang J, Yang D, Deng R, Wan Y, Yan B (2006) The expression of antiapoptotic protein survivin is transcriptionally upregulated by DEC1 primarily through multiple sp1 binding sites in the proximal promoter. Oncogene 25:3296–3306

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  38. Cosgrave N, Hill AD, Young LS (2006) Growth factor-dependent regulation of survivin by c-myc in human breast cancer. J Mol Endocrinol 37:377–390

    Article  CAS  PubMed  Google Scholar 

  39. Zhu N, Gu L, Findley HW, Chen C, Dong JT, Yang L, Zhou M (2006) KLF5 Interacts with p53 in regulating survivin expression in acute lymphoblastic leukemia. J Biol Chem 281:14711–14718

    Article  CAS  PubMed  Google Scholar 

  40. Gritsko T, Williams A, Turkson J, Kaneko S, Bowman T, Huang M, Nam S, Eweis I, Diaz N, Sullivan D, Yoder S, Enkemann S, Eschrich S, Lee JH, Beam CA, Cheng J, Minton S, Muro-Cacho CA, Jove R (2006) Persistent activation of stat3 signaling induces survivin gene expression and confers resistance to apoptosis in human breast cancer cells. Clin Cancer Res 12:11–19

    Article  CAS  PubMed  Google Scholar 

  41. Peng XH, Karna P, Cao Z, Jiang BH, Zhou M, Yang L (2006) Cross-talk between epidermal growth factor receptor and hypoxia-inducible factor-1alpha signal pathways increases resistance to apoptosis by up-regulating survivin gene expression. J Biol Chem 281:25903–25914

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  42. Dasgupta P, Kinkade R, Joshi B, Decook C, Haura E, Chellappan S (2006) Nicotine inhibits apoptosis induced by chemotherapeutic drugs by up-regulating XIAP and survivin. Proc Natl Acad Sci USA 103:6332–6337

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  43. Kim YH, Lee DH, Jeong JH, Guo ZS, Lee YJ (2008) Quercetin augments TRAIL-induced apoptotic death: involvement of the ERK signal transduction pathway. Biochem Pharmacol 75:1946–1958

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  44. Ringshausen I, Schneller F, Bogner C, Hipp S, Duyster J, Peschel C, Decker T (2002) Constitutively activated phosphatidylinositol-3 kinase (PI-3 K) is involved in the defect of apoptosis in B-CLL: association with protein kinase Cdelta. Blood 100:3741–3748

    Article  CAS  PubMed  Google Scholar 

  45. Hu P, Han Z, Couvillon AD, Exton JH (2004) Critical role of endogenous Akt/IAPs and MEK1/ERK pathways in counteracting endoplasmic reticulum stress-induced cell death. J Biol Chem 279:49420–49429

    Article  CAS  PubMed  Google Scholar 

  46. Abdelrahim M, Liu S, Safe S (2005) Induction of endoplasmic reticulum-induced stress genes in Panc-1 pancreatic cancer cells is dependent on Sp proteins. J Biol Chem 280:16508–16513

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by Basic Science Research Program through the NRF funded by the Ministry of Education, Science and Technology (2010-0009064).

Author information

Authors and Affiliations

  1. Department of Molecular Biology and Immunology, Kosin University College of Medicine, 265 Gamcheon-ro, Seo-gu, Busan, 602-702, Korea

    Eun Jin Lim, Jeonghoon Heo & Young-Ho Kim

  2. Institute for Medical Sciences, Kosin University College of Medicine, Busan, Korea

    Jeonghoon Heo & Young-Ho Kim

Authors
  1. Eun Jin Lim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jeonghoon Heo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Young-Ho Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Young-Ho Kim.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lim, E.J., Heo, J. & Kim, YH. Tunicamycin promotes apoptosis in leukemia cells through ROS generation and downregulation of survivin expression. Apoptosis 20, 1087–1098 (2015). https://doi.org/10.1007/s10495-015-1135-z

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10495-015-1135-z

Keywords

Search

Navigation