Veterinary Research Forum
Vet Res Forum

Cytotoxic effect of sea anemone pore-forming toxin on K562 chronic myeloid leukemia cells

Document Type : Original Article

Authors

Vali Abdoli 1
Roya Sarkhosh-Inanlou 2
Nowruz Delirezh 2, 3
Safiyeh Aghazadeh 1
Nima ShaykhBaygloo 4
Mehdi Imani 1, 2

1 Department of Basic Science, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

2 Department of Cellular and Molecular Biotechnology, Institute of Biotechnology, Urmia University, Urmia, Iran

3 Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

4 Department of Biology, Faculty of Sciences, Urmia University, Urmia, Iran

https://doi.org/10.30466/vrf.2020.115033.2737
Abstract
Chronic myelogenous leukemia (CML) is one of prevalent cancer worldwide. In spite of various designed drugs, chemoresistance remains the main obstacle in cancer cure. Therefore, developing novel strategy for treatment of CML is an urgent need. Fragaceatoxin C (FraC) is novel protein toxin from a sea anemone called actinia fragacea with great impacts against cells by pore formation and disturbing cell membrane integrity. The aim of this study was evaluation of FraC toxin toxicity against K562. The bacteria cells harboring expression‌‌‌‌‌‌‌ vector of FraC were induced by IPTG and purified by Ni2+-NTA sepharose affinity chromatography. Then, purified toxin activity was evaluated using RBC hemolytic test. Eventually, evaluation of FraC cytotoxicity and apoptosis were performed using MTT and flow cytometery assays, respectively. Our results revealed that FraC toxin decreased K562 cells viability in a dose- and time-dependent manner with a whole destroy of cancer cells at 35.00 µg mL-1 after 72 hr. Furthermore, flow cytometery analysis indicated that FraC toxin enhanced necrosis along with apoptosis in K562 cells in a dose dependent manner. We speculated that FraC toxin could be considered as a novel candidate for cancer cell researches and treatments provided that it should be turned into a specific agent by engineering and directing to cancer cell membrane.

Keywords

Chronic myelogenous leukemia
Fragaceatoxin C
Necrosis
Pore-forming toxin
  1. Mirzaei H, Fathullahzadeh S, Khanmohammadi R, et al. State of the art in microRNA as diagnostic and therapeutic biomarkers in chronic lymphocytic leukemia. J Cell Physiol 2018; 233(2): 888-900.
  2. Fathullahzadeh S, Mirzaei H, Honardoost MA, et al. Circulating microRNA-192 as a diagnostic biomarker in human chronic lymphocytic leukemia. Cancer Gene Ther 2016; 23(10): 327-332.
  3. Brigger I, Dubernet C, Couvreur P. Nanoparticles in cancer therapy and diagnosis. Adv Drug Deliv Rev 2002; 54(5): 631-651.
  4. Di Francia R, De Monaco A, Saggese M, et al. Pharmacological profile and pharmacogenomics of anti-cancer drugs used for targeted therapy. Curr Cancer Drug Targets 2018; 18(5): 499-511.
  5. Saeidnia S, Abdollahi M. Perspective studies on novel anticancer drugs from natural origin: a comprehensive review. Int J Pharmacol 2014; 10(2): 90-108.
  6. Mutter NL, Soskine M, Huang G, et al. Modular pore-forming immunotoxins with caged cytotoxicity tailored by directed evolution. ACS Chem Biol 2018; 13(11): 3153-3160.
  7. Azadpour M, Karimian M, Kheirandish MH, et al. Evaluating cytotoxic effects of recombinant fragaceatoxin C pore forming toxin against AML cell lines. Iran J Basic Med Sci 2018; 21(9): 878-883.
  8. Imani M, Jaliani HZ, Kheirandish MH et al. Recombinant production and affinity purification of the FraC pore forming toxin using hexa-His tag and pET expression cassette. Iran J Basic Med Sci 2017; 20(4): 380-385.
  9. DuMont AL, Torres VJ. Cell targeting by the Staphylococcus aureus pore-forming toxins: it's not just about lipids. Trends Microbiol 2014; 22(1): 21-27.
  10. Patyar S, Joshi R, Byrav DS, et al. Bacteria in cancer therapy: a novel experimental strategy. J Biomed Sci 2010; 17(1): 21. doi: 10.1186/1423-0127-17-21.
  11. Soletti RC, de Faria GP, Vernal J, et al. Potentiation of anticancer-drug cytotoxicity by sea anemone pore-forming proteins in human glioblastoma cells. Anticancer Drugs 2008; 19(5): 517-525.
  12. Perrin MC, Thorpe JP, Solé-Cava AM. Population structuring, gene dispersal and reproduction in the Actinia equina species group. In: Ansell A, Gibson RN, Barnes M (Eds). Oceanography and marine biology, an annual review. Volume 37. 1st London, UK: CRC Press 1999; 129-152.
  13. Bellomio A, Morante K, Barlic A, et al. Purification, cloning and characterization of fragaceatoxin C, a novel actinoporin from the sea anemone Actinia fragacea. Toxicon 2009; 54(6): 869-880.
  14. Morante K, Bellomio A, Viguera AR, et al. The isolation of new pore-forming toxins from the sea anemone Actinia fragacea provides insights into the mechanisms of actinoporin evolution. Toxins (Basel) 2019; 11(7): 401. doi: 10.3390/toxins11070401.
  15. Mechaly AE, Bellomio A, Gil-Cartón D, et al. Structural insights into the oligomerization and architecture of eukaryotic membrane pore-forming toxins. Structure 2011; 19(2): 181-191.
  16. Yang WS, Park SO, Yoon AR, et al. Suicide cancer gene therapy using pore-forming toxin, streptolysin O. Mol Cancer Ther 2006; 5(6): 1610-1619.
  17. Ravindran VS, Kannan L, Venkateshvaran K. Biological activity of sea anemone proteins: II. Cytolysis and cell line toxicity. Indian J Exp Biol 2010; 48(12):1233-1236
  18. Bulati M, Longo A, Masullo T, et al. Partially purified extracts of sea anemone Anemonia viridis affect the growth and viability of selected tumour cell lines. Biomed Res Int 2016; 3849897. doi:10.1155/2016/3849897.
  19. García-Linares S, Alm I, Maula T, et al. The effect of cholesterol on the long-range network of interactions established among sea anemone Sticholysin II residues at the water-membrane interface. Mar Drugs 2015; 13(4): 1647-1665.
  20. Kramer MG, Masner M, Ferreira FA, et al. Bacterial therapy of cancer: promises, limitations, and insights for future directions. Front Microbiol 2018; 9: 16. doi: 10.3389/fmicb.2018.00016.
  21. Kaimala S, Al-Sbiei A, Cabral-Marques O, et al. Attenuated bacteria as immunotherapeutic tools for cancer treatment. Front Oncol 2018; 8: 136. doi: 10.3389/fonc.2018.00136.
  22. Rea D, Coppola G, Palma G, et al. Microbiota effects on cancer: from risks to therapies. Oncotarget 2018; 9(25): 17915-17927.
  23. Pahle J, Walther W. Bacterial toxins for oncoleaking suicidal cancer gene therapy. Recent Results Cancer Res 2016; 209: 95-110.
Veterinary Research Forum
Volume 12, Issue 4
Autumn 2021
Pages 481-485

  • Receive Date 29 September 2019
  • Revise Date 07 April 2020
  • Accept Date 08 April 2020

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