Tumor Biology

, Volume 37, Issue 10, pp 14291–14300 | Cite as

Armillaridin induces autophagy-associated cell death in human chronic myelogenous leukemia K562 cells

Original Article

Abstract

Armillaridin (AM) is an aromatic ester compound isolated from Armillaria mellea. Treatment with AM markedly reduced the viability of human chronic myelogenous leukemia K562, chronic erythroleukemia HEL 92.1.7, and acute monoblastic leukemia U937 cells, but not normal human monocytes, in a dose- and time-dependent manner. Treatment of K562 cells with AM caused changes characteristic of autophagy. Only a small amount of AM-treated K562 cells exhibited apoptosis. By contrast, AM treatment resulted in extensive apoptotic features in U937 and HEL 92.1.7 cells without evident autophagy. The autophagy of K562 cells induced by AM involved autophagic flux, including autophagosome induction, the processing of autophagosome-lysosome fusion and downregulation of BCL2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3). By bcr-abl knockdown, the growth inhibition of K562 cells caused by AM was partially blocked, suggesting that AM-induced cell death might be a bcr-abl-dependent mode of autophagy-associated cell death. In conclusion, AM is capable of inhibiting growth and inducing autophagy-associated cell death in K562 cells, but not in normal monocytes. It may have potential to be developed as a novel therapeutic agent against leukemia.

Keywords

Armillaridin Autophagy bcr-abl BNIP3 Chronic myelogenous leukemia 

Abbreviations

AM

Armillaridin

BNIP3

BCL2/adenovirus E1B 19 kDa interacting protein 3

CML

Chronic myelogenous leukemia

CQ

Chloroquine

3-MA

3-Methyladenine

STI-571

Signal transduction inhibitor-571

Notes

Acknowledgments

We want to thank Dr. King-Song Jeng and the National RNAi Core Facility, Academia Sinica, Taiwan, for shRNA knockdown techniques, the pTRC-905 vector, and the pTRC-905-shBCR constructions. This work was supported by grant NSC98-2323-B-241 from the National Science Council, Taiwan, and grant 09MMHIS027 and MMH-E-105-13, E-104-13 from MacKay Memorial Hospital, Taiwan.

Compliance with ethical standards

Conflicts of interest

None.

Supplementary material

13277_2016_5208_Fig10_ESM.gif (176 kb)
Supplemental Figure 1.

Effect of armillaridin on BCR-ABL expression on K562 cells. Cells were untreated, treated with 0.1% DMSO (solvent), 10 μM armillaridin for 24 hours or starvation for 6 hours. Protein extracts from whole cells were subjected to Western blotting. Data from three separate experiments showed similar trend. No significant differences on the expression of BCR-ABL were found between control and armillaridin treatment. (GIF 175 kb)

13277_2016_5208_MOESM1_ESM.tiff (1.5 mb)
High Resolution Image (TIFF 1521 kb)

References

  1. 1.
    Hehlmann R, Hochhaus A, Baccarani M, European L. Chronic myeloid leukaemia. Lancet. 2007;370:342–50.CrossRefPubMedGoogle Scholar
  2. 2.
    Apperley JF. Chronic myeloid leukaemia. Lancet. 2014.Google Scholar
  3. 3.
    Bellodi C, Lidonnici MR, Hamilton A, Helgason GV, Soliera AR, Ronchetti M, Galavotti S, Young KW, Selmi T, Yacobi R, Van Etten RA, Donato N, Hunter A, Dinsdale D, Tirro E, Vigneri P, Nicotera P, Dyer MJ, Holyoake T, Salomoni P, Calabretta B. Targeting autophagy potentiates tyrosine kinase inhibitor-induced cell death in Philadelphia chromosome-positive cells, including primary CML stem cells. J Clin Invest. 2009;119:1109–23.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Lung MY, Chang YC. Antioxidant properties of the edible Basidiomycete Armillaria mellea in submerged cultures. Int J Mol Sci. 2011;12:6367–84.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Yang JS, Chen YW, Feng XZ, Yu DQ, Liang XT. Chemical constituents of Armillaria mellea mycelium. I. Isolation and characterization of armillarin and armillaridin. Planta Med. 1984;50:288–90.CrossRefPubMedGoogle Scholar
  6. 6.
    Chi CW, Chen CC, Chen YJ. Therapeutic and radiosensitizing effects of armillaridin on human esophageal cancer cells. Evidence-based complementary and alternative medicine: eCAM. 2013;2013:459271.Google Scholar
  7. 7.
    Glick D, Barth S, Macleod KF. Autophagy: cellular and molecular mechanisms. J Pathol. 2010;221:3–12.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Marino G, Niso-Santano M, Baehrecke EH, Kroemer G. Self-consumption: the interplay of autophagy and apoptosis. Nat Rev Mol Cell Biol. 2014;15:81–94.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Strober W. Trypan blue exclusion test of cell viability. Current protocols in immunology/edited by John E Coligan [et al]. 2001;Appendix 3:Appendix 3B.Google Scholar
  10. 10.
    Puissant A, Grosso S, Jacquel A, Belhacene N, Colosetti P, Cassuto JP, Auberger P. Imatinib mesylate-resistant human chronic myelogenous leukemia cell lines exhibit high sensitivity to the phytoalexin resveratrol. FASEB J. 2008;22:1894–904.CrossRefPubMedGoogle Scholar
  11. 11.
    Sen N, Das BB, Ganguly A, Mukherjee T, Tripathi G, Bandyopadhyay S, Rakshit S, Sen T, Majumder HK. Camptothecin induced mitochondrial dysfunction leading to programmed cell death in unicellular hemoflagellate Leishmania donovani. Cell Death Differ. 2004;11:924–36.CrossRefPubMedGoogle Scholar
  12. 12.
    Huang HL, Chen YC, Huang YC, Yang KC, Pan H, Shih SP, Chen YJ. Lapatinib induces autophagy, apoptosis and megakaryocytic differentiation in chronic myelogenous leukemia K562 cells. PLoS One. 2011;6:e29014.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Schneider CA, Rasband WS, Eliceiri KW. NIH image to ImageJ: 25 years of image analysis. Nat Methods. 2012;9:671–5.CrossRefPubMedGoogle Scholar
  14. 14.
    Jacquel A, Herrant M, Legros L, Belhacene N, Luciano F, Pages G, Hofman P, Auberger P. Imatinib induces mitochondria-dependent apoptosis of the Bcr-Abl-positive K562 cell line and its differentiation toward the erythroid lineage. FASEB J. 2003;17:2160–2.CrossRefPubMedGoogle Scholar
  15. 15.
    Azad MB, Gibson SB. Role of BNIP3 in proliferation and hypoxia-induced autophagy: implications for personalized cancer therapies. Ann N Y Acad Sci. 2010;1210:8–16.CrossRefPubMedGoogle Scholar
  16. 16.
    He C, Klionsky DJ. Regulation mechanisms and signaling pathways of autophagy. Annu Rev Genet. 2009;43:67–93.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Daido S, Kanzawa T, Yamamoto A, Takeuchi H, Kondo Y, Kondo S. Pivotal role of the cell death factor BNIP3 in ceramide-induced autophagic cell death in malignant glioma cells. Cancer Res. 2004;64:4286–93.CrossRefPubMedGoogle Scholar
  18. 18.
    Chen YJ, Huang WP, Yang YC, Lin CP, Chen SH, Hsu ML, Tseng YJ, Shieh HR, Chen YY, Lee JJ. Platonin induces autophagy-associated cell death in human leukemia cells. Autophagy. 2009;5:173–83.CrossRefPubMedGoogle Scholar
  19. 19.
    Kanzawa T, Zhang L, Xiao L, Germano IM, Kondo Y, Kondo S. Arsenic trioxide induces autophagic cell death in malignant glioma cells by upregulation of mitochondrial cell death protein bnip3. Oncogene. 2005;24:980–91.CrossRefPubMedGoogle Scholar
  20. 20.
    Cheng Y, Qiu F, Huang J, Tashiro S, Onodera S, Ikejima T. Apoptosis-suppressing and autophagy-promoting effects of calpain on oridonin-induced L929 cell death. Arch Biochem Biophys. 2008;475:148–55.CrossRefPubMedGoogle Scholar
  21. 21.
    Goussetis DJ, Gounaris E, EJ W, Vakana E, Sharma B, Bogyo M, Altman JK, Platanias LC. Autophagic degradation of the BCR-ABL oncoprotein and generation of antileukemic responses by arsenic trioxide. Blood. 2012;120:3555–62.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Sheng Z, Ma L, Sun JE, Zhu LJ, Green MR. BCR-ABL suppresses autophagy through ATF5-mediated regulation of mTOR transcription. Blood. 2011;118:2840–8.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  1. 1.Institute of Traditional MedicineNational Yang-Ming UniversityTaipeiTaiwan
  2. 2.Department of Healthcare CenterMacKay Memorial HospitalNew Taipei CityTaiwan
  3. 3.Department of Bioscience Technology, College of Health ScienceChang Jung Christian UniversityTainanTaiwan
  4. 4.Department of Life Science and Institute of ZoologyNational Taiwan UniversityTaipeiTaiwan
  5. 5.Department of BiotechnologyHungkuang UniversityTaichung CityTaiwan
  6. 6.Department of Medical ResearchMacKay Memorial HospitalTaipei CityTaiwan
  7. 7.Department of Radiation OncologyMacKay Memorial HospitalTaipei CityTaiwan

Personalised recommendations