Document Type : Original Article

Authors

1 Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

2 Department of Basic Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

3 Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

4 Department of Clinical Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

Abstract

Given the development of drug-resistant cancer cells, designing alternative approaches for cancer treatment seems essential. In this study, we evaluated the anti-tumor effects of nisin A and newcastle disease virus (NDV) on triple-negative MDA-MB-231 cell line. The MDA-MB-231 cell line was separately and in combination subjected to the different concentrations of a Vero-adapted NDV (JF820294.1) and nisin A. The oncolytic effects of these treatments were analyzed by different cytotoxic and apoptosis techniques including trypan blue staining, MTT assay, acridine orange (EB/AO) staining, colony assay and flow cytometry over time. Nisin A at doses of more than 20.00 μg mL-1 could represent the anti-viral effects and interfere with the oncolytic activity of NDV. Moreover, the analyses indicated that the anti-proliferative and cytotoxic features of combination therapy were stronger than those of individual NDV groups. However, the most apoptotic effect was seen in NDV experimental groups. Taken together, the results from cytotoxicity tests, flow cytometry and colony assay showed that either of the oncolytic agents had significant effects at low concentrations 72 hr post-treatment. Thereby, they had the potential to be used as new approaches in cancer treatment.

Keywords

  1. Anastasiadi Z, Lianos GD, Ignatiadou E, et al. Breast cancer in young women: an overview. Updates Surg 2017; 69(3): 313-317.
  2. Bodai BI, Tuso P. Breast cancer survivorship: a comprehensive review of long-term medical issues and lifestyle recommendations. Perm J 2015; 19(2): 48-79.
  3. Bourgeois-Daigneault MC, St-Germain LE, Roy DG, et al. Combination of Paclitaxel and MG1 oncolytic virus as a successful strategy for breast cancer treatment. Breast Cancer Res 2016; 18: 83. doi: 10.1186/s13058-016-0744-y.
  4. Ranaware AM, Banik K, Deshpande V, et al. Magnolol: a neolignan from the magnolia family for the prevention and treatment of cancer. Int J Mol Sci 2018; 19(8): 2362. doi: 10.3390/ijms19082362.
  5. Wang X, Yang Y, An Y, et al. The mechanism of anticancer action and potential clinical use of kaempferol in the treatment of breast cancer. Biomed Pharmacother 2019; 117: 109086. doi: 10.1016/ j.biopha.2019.109086.
  6. Zeng F, Ju RJ, Liu L, et al. Efficacy in treating lung metastasis of invasive breast cancer with functional vincristine plus dasatinib liposomes. Pharmacology 2018; 101(1-2): 43-53.
  7. Hosseini SS, Aghaiypour Kolyani K, Rafiei Tabatabaei R, et al. In silico prediction of B and T cell epitopes based on NDV fusion protein for vaccine development against Newcastle disease virus. Vet Res Forum 2021; 12(2): 157-165.
  8. Mehrabanpour MJ, Khoobyar S, Rahimian A, et al. Phylogenetic characterization of the fusion genes of the Newcastle disease viruses isolated in Fars province poultry farms during 2009-2011. Vet Res Forum 2014; 5(3): 187-191.
  9. Lam HY, Yeap SK, Pirozyan MR, et al. Corrigendum to "safety and clinical usage of Newcastle disease virus in cancer therapy". Biomed Res Int 2017; 2017: 4529437. doi: 10.1155/2017/4529437.
  10. Prince AM, Ginsberg HS. Immunohistochemical studies on the interaction between Ehrlich ascites tumor cells and Newcastle disease virus. J Exp Med 1957; 105(2): 177-188.
  11. Eaton MD, Levinthal JD, Scala AR. Contribution of antiviral immunity to oncolysis by Newcastle disease virus in a murine lymphoma. J Natl Cancer Inst 1967; 39(6): 1089-1097.
  12. Shi J, Sun P, Zhang Y, et al. The antitumor effects of Newcastle disease virus on glioma. Biocell 2019; 43(3): 119-128.
  13. Buijs P, van Nieuwkoop S, Vaes V, et al. Recombinant immunomodulating lentogenic or mesogenic oncolytic Newcastle disease virus for treatment of pancreatic adenocarcinoma. Viruses 2015; 7(6): 2980-2998.
  14. Zamarin D, Martínez-Sobrido L, Kelly K, et al. Enhancement of oncolytic properties of recombinant Newcastle disease virus through antagonism of cellular innate immune responses. Mol Ther 2009; 17(4): 697-706.
  15. Beyzaei H, Mirzaei M, Fakhrabadi N, et al. Synergistic effects of dual antimicrobial combinations of synthesized N-heterocycles or MgO nanoparticles with nisin against the growth of Aspergillus fumigatus: In vitro Vet Res Forum 2021; 12(2): 241-246.
  16. Shahbazi Y. The antibacterial effect of Ziziphora clinopodioides essential oil and nisin against Salmonella typhimurium and Staphylococcus aureus in doogh, a yoghurt-based Iranian drink. Vet Res Forum 2016; 7(3): 213-219.
  17. World Health Organization website. Specifications for the identity and purity of food additives and their toxicological evaluation: Some antibiotics. Available at: https://oem.bmj.com/content/27/2/185.3. Accessed 22 Nov 2022.
  18. Food and Drug Administration. Nisin preparation: affirmation of GRAS status as a direct human food ingredient. Available at https://www.scribd.com/doc-ument/312619717/Nisin-Preparation-Affirmation-of-GRAS-Status-as-a-Direct-Human-Food-Ingredient-FAO. Accessed 22 Nov 2022.
  19. Ahmadi S, Ghollasi M, Hosseini HM. The apoptotic impact of nisin as a potent bacteriocin on the colon cancer cells. Microb Pathog 2017; 111: 193-197.
  20. Avand A, Akbari V, Shafizadegan S. In vitro cytotoxic activity of a Lactococcus lactis antimicrobial peptide against breast cancer cells. Iran J Biotechnol 2018; 16(3): e1867. doi: 10.15171/ijb.1867.
  21. Kamarajan P, Hayami T, Matte B, et al. Nisin ZP, a bacteriocin and food preservative, inhibits head and neck cancer tumorigenesis and prolongs survival. PloS One 2015; 10(7): e0131008. doi: 10.1371/journal. pone.0131008.
  22. Preet S, Bharati S, Panjeta A, et al. Effect of nisin and doxorubicin on DMBA-induced skin carcinogenesis--a possible adjunct therapy. Tumour Biol 2015; 36(11): 8301-8308.
  23. Zainodini N, Hassanshahi G, Hajizadeh M, et al. Nisin induces cytotoxicity and apoptosis in human asterocytoma cell line (SW1088). Asian Pac J Cancer Prev 2018; 19(8): 2217-2222.
  24. Otvos L Jr. Immunomodulatory effects of anti-microbial peptides. Acta Microbiol Immunol Hung 2016; 63(3): 257-277.
  25. Rodrigues G, Silva GGO, Buccini DF, et al. Bacterial proteinaceous compounds with multiple activities toward cancers and microbial infection. Front Microbiol 2019; 10: 1690. doi: 10.3389/fmicb. 2019.01690.
  26. Shin JM, Gwak JW, Kamarajan P, et al. Biomedical applications of nisin. J Appl Microbiol 2016; 120(6): 1449-1465.
  27. Moein M, Imani Fooladi AA, Mahmoodzadeh Hosseini H. Determining the effects of green chemistry synthesized Ag-nisin nanoparticle on macrophage cells. Microb Pathog 2018; 114: 414-419.
  28. Goudarzi F, Asadi A, Afsharpour M, et al. In vitro characterization and evaluation of the cytotoxicity effects of nisin and nisin-loaded PLA-PEG-PLA nanoparticles on gastrointestinal (AGS and KYSE-30), hepatic (HepG2) and blood (K562) cancer cell lines. AAPS PharmSciTech 2018; 19(4): 1554-1566.
  29. Adusumilli PS, Stiles BM, Chan MK, et al. Radiation therapy potentiates effective oncolytic viral therapy in the treatment of lung cancer. Ann Thorac Surg 2005; 80(2): 409-416.
  30. Bourgeois-Daigneault MC, Roy DG, Aitken AS, et al. Neoadjuvant oncolytic virotherapy before surgery sensitizes triple-negative breast cancer to immune checkpoint therapy. Sci Transl Med 2018; 10(422): eaao1641. doi: 10.1126/scitranslmed.aao1641.
  31. Ilett E, Kottke T, Thompson J, et al. Prime-boost using separate oncolytic viruses in combination with checkpoint blockade improves anti-tumour therapy. Gene Ther 2017; 24(1): 21-30.
  32. Khuri FR, Nemunaitis J, Ganly I, et al. A controlled trial of intratumoral ONYX-015, a selectively-replicating adenovirus, in combination with cisplatin and 5-fluorouracil in patients with recurrent head and neck cancer. Nat Med 2000; 6(8): 879-885.
  33. Kleinpeter P, Fend L, Thioudellet C, et al. Vectorization in an oncolytic vaccinia virus of an antibody, a Fab and a scFv against programmed cell death-1 (PD-1) allows their intratumoral delivery and an improved tumor-growth inhibition. Oncoimmunology 2016; 5(10): e1220467. doi: 10.1080/2162402X.2016.1220467.
  34. Nishio N, Dotti G. Oncolytic virus expressing RANTES and IL-15 enhances function of CAR-modified T cells in solid tumors. Oncoimmunology 2015; 4(2): e988098. doi: 10.4161/21505594.2014.988098.
  35. O’Cathail SM, Pokrovska TD, Maughan TS, et al. Combining oncolytic adenovirus with radiation-a paradigm for the future of radiosensitization. Front Oncol 2017; 7: 153. doi: 10.3389/fonc.2017.00153.
  36. Puzanov I, Milhem MM, Minor D, et al. Talimogene laherparepvec in combination with ipilimumab in previously untreated, unresectable stage IIIB-IV melanoma. J Clin Oncol 2016; 34(22): 2619-2626.
  37. Roulstone V, Pedersen M, Kyula J, et al. BRAF-and MEK-targeted small molecule inhibitors exert enhanced antimelanoma effects in combination with oncolytic reovirus through ER stress. Mol Ther 2015; 23(5): 931-942.
  38. Silk A, Kaufman H, Gabrail N, et al. Abstract CT026: Phase 1b study of intratumoral Coxsackievirus A21 (CVA21) and systemic pembrolizumab in advanced melanoma patients: Interim results of the CAPRA clinical trial. Cancer Res 2017; 77(13 Supplement): CT026- doi: 10.1158/1538-7445.AM2017-CT026.
  39. Zamarin D, Holmgaard RB, Subudhi SK, et al. Localized oncolytic virotherapy overcomes systemic tumor resistance to immune checkpoint blockade immunotherapy. Sci Transl Med 2014; 6(226): 226ra32. doi: 10.1126/scitranslmed.3008095.
  40. Bommareddy PK, Shettigar M, Kaufman HL. Integrating oncolytic viruses in combination cancer immuno-therapy. Nat Rev Immunol 2018; 18(8): 498-513.
  41. Habibi H, Nili H, Asasi K, et al. Efficacy and transmissibility of Newcastle disease I-2 vaccine strain against a field isolate of virulent ND virus (JF820294.1) in village chicken. Trop Anim Health Prod 2015; 47(1): 73-78.
  42. Ribble D, Goldstein NB, Norris DA, et al. A simple technique for quantifying apoptosis in 96-well plates. BMC Biotechnol 2005; 5: 12. doi: 10.1186/1472-6750-5-12.
  43. Cohen JJ. Apoptosis. Immunol Today 1993; 14(3): 126-130.
  44. Małaczewska J, Kaczorek-Łukowska E, Wójcik R, et al. Antiviral effects of nisin, lysozyme, lactoferrin and their mixtures against bovine viral diarrhoea virus. BMC Vet Res 2019; 15(1): 318. doi: 10.1186/s12917-019-2067-6.
  45. Chung DM, Kim JH, Kim JK. Evaluation of MTT and Trypan Blue assays for radiation-induced cell viability test in HepG2 cells. Int J Radiat Res 2015; 13(4): 331-335.
  46. Braselmann H, Michna A, Heß J, et al. CFAssay: statistical analysis of the colony formation assay. Radiat Oncol 2015; 10: 223. doi: 10.1186/s13014-015-0529-y.