Isolation and identification of the bacterium producing antitumor and antimicrobial compounds derived from Iranian swamp frog (Rana ridibunda) skin
,
Abstract
Background and Objectives: Cancer incidence and recurrence, antibiotic resistance, and overuse of antibiotics have become a global concern. The purpose of this study was to identify and isolate bacteria from the skin of the Rana ridibunda, Iranian swamp frog, which has produced antimicrobial compounds, and investigate its cytotoxic activity on the breast (MCF7) and glioblastoma (U87) cancer cell line.
Materials and Methods: An antibiotic-producing bacterium was isolated from the frog skin. The bacterium was identified based on 16S rDNA sequencing and biochemical and morphological characteristics. Antimicrobial activity of the culture supernatant was examined by disc diffusion and MIC methods. Cytoplasmic and cell wall extracts of bacteria were prepared by sonication. SDS-PAGE was then used to examine protein contents of them. The cancer cell lines were treated with cytoplasmic and cell wall extracts at different concentrations. The effects of cytotoxicity were assessed by MTT assay at 24 and 48 h intervals. Finally, the results were analyzed by SPSS.
Results: The isolated bacterium was identified as a new strain of Bacillus atrophaeus. MIC and disc diffusion methods showed that the Bacillus atrophaeus antimicrobial activity was broad spectrum. MTT assay showed IC50 values 30 μg/ml and 20 μg/ml for U87 and MCF7 cells after 24-48 h exposure, respectively.
Conclusion: The cytoplasmic extracts of Bacillus atrophaeus has anticancer potential and can be used as an alternative or complementary candidate in the treatment of cancer. Further in vivo and in vitro mechanistic studies are suggested to confirm the biological activities.
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24. Thon N, Thorsteinsdottir J, Eigenbrod S, Schüller U, Lutz J, Kreth S, et al. Outcome in unresectable glioblastoma: MGMT promoter methylation makes the difference. J Neurol 2017;264:350-358.
25. Scatena C, Roncella M, Di Paolo A, Aretini P, Menicagli M, Fanelli G, et al. Doxycycline, an inhibitor of mitochondrial biogenesis, effectively reduces cancer stem cells (CSCs) in early breast cancer patients: a clinical pilot study. Front Oncol 2018;8:452.
26. Lamb R, Fiorillo M, Chadwick A, Ozsvari B, Reeves KJ, Smith DL, et al. Doxycycline down-regulates DNA-PK and radiosensitizes tumor initiating cells: implications for more effective radiation therapy. Oncotarget 2015;6:14005-14025.
27. Jalali F, Sharifi M, Salehi R. Kefir induces apoptosis and inhibits cell proliferation in human acute erythroleukemia. Med Oncol 2016;33:7.
28. Rizk S, Maalouf K, Baydoun E. The antiproliferative effect of kefir cell-free fraction on HuT-102 malignant T lymphocytes. Clin Lymphoma Myeloma 2009;9 Suppl 3:S198-203.
29. Vidhyalakshmi R, Vallinachiyar C. Apoptosis of human breast cancer cells (MCF-7) induced by polysacccharides produced by bacteria. J Cancer Sci Ther 2013;5:31-34.
30. Abdulrahman SA, Abou El-E GA, Gamal HO. Overexpression and cytocidal activity of parasporin-1 from Bacillus thuringiensis against human cancer cell lines. J Microb Biochem Technol 2017;9:220-226.
31. St Jean AT, Swofford CA, Panteli JT, Brentzel ZJ, Forbes NS. Bacterial delivery of Staphylococcus aureus α-hemolysin causes regression and necrosis in murine tumors. Mol Ther 2014;22:1266-1274.
32. Kumar MV, Thippeswamy B, Raj PV. Cytotoxicity and anticancer studies of Bacillus cereus and Bacillus pumilus metabolites targeting human cancer cells. Appl Biochem Microbiol 2014;50:619-623.
33. Cao X, Wang AH, Jiao RZ, Wang CL, Mao DZ, Yan L, et al. Surfactin induces apoptosis and G (2)/M arrest in human breast cancer MCF-7 cells through cell cycle factor regulation. Cell Biochem Biophys 2009;55:163-171.
34. Aldeewan A, Zhang Y, Su L. Bacillus thuringiensis parasporins functions on cancer cells. Int J Pure Appl Biosci 2014;2:67-74.
2. Anampa J, Sparano JA. New agents for the management of resistant metastatic breast cancer. Expert Opin Pharmacother 2017;18:1815-1831.
3. Cheetham PJ, Petrylak DP. New agents for the treatment of advanced bladder cancer. Oncology (Williston Park) 2016;30:571-579,588.
4. Stewart PS, Costerton JW. Antibiotic resistance of bacteria in biofilms. Lancet 2001;358:135-138.
5. Hancock RE, Scott MG. The role of antimicrobial peptides in animal defenses. Proc Natl Acad Sci U S A 2000;97:8856-8861.
6. Zasloff M. Antimicrobial peptides of multicellular organisms. Nature 2002;415:389-395.
7. Rivas L, Luque-Ortega JR, Andreu D. Amphibian antimicrobial peptides and Protozoa: lessons from parasites. Biochim Biophys Acta 2009;1788:1570-1581.
8. Temmerman R, Pot B, Huys G, Swings J. Identification and antibiotic susceptibility of bacterial isolates from probiotic products. Int J Food Microbiol 2003;81:1-10.
9. Adiguzel A, Ozkan H, Baris O, Inan K, Gulluce M, Sahin F. Identification and characterization of thermophilic bacteria isolated from hot springs in Turkey. J Microbiol Methods 2009;79:321-328.
10. Ebrahimipour GH, Khosravibabadi Z, Sadeghi H, Aliahmadi A. Isolation, partial purification and characterization of an antimicrobial compound, produced by Bacillus atrophaeus. Jundishapur J Microbiol 2014;7(9):e11802.
11. Mardis ER. DNA sequencing technologies: 2006–2016. Nat Protoc 2017;12: 213-218.
12. Dworkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E (2006). The prokaryotes: vol. 4: bacteria: firmicutes, cyanobacteria. Springer Science & Business Media. 3rd ed. New York, NY: Springer.
13. Beiranvand M, Amin M, Hashemi-Shahraki A, Romani B, Yaghoubi S, Sadeghi P. Antimicrobial activity of endophytic bacterial populations isolated from medical plants of Iran. Iran J Microbiol 2017;9:11-18.
14. Lahuerta Zamora L, Perez-Gracia MT. Using digital photography to implement the McFarland method. J R Soc Interface 2012;9:1892-1897.
15. Zapata A, Ramirez-Arcos S. A comparative study of McFarland turbidity standards and the Densimat photometer to determine bacterial cell density. Curr Microbiol 2015;70:907-909.
16. Fatahi A, Soleimani N. Evaluation of cytotoxicity activity of cell extracts of kefir microorganisms on glioblastoma cancer cells. Stud Med Sci 2018;29:12-19.
17. Soleimani N, Farhangi B, Tavakoli Yaraki M. The effect of recombinant HopH protein of Helicobacter pylori on the VEGF expression in metastatic breast cancer model. Acta Med Iran 2017;55:744-750.
18. Yousofi A, Daneshmandi S, Soleimani N, Bagheri K, Karimi MH. Immunomodulatory effect of Parsley (Petroselinum crispum) essential oil on immune cells: mitogen-activated splenocytes and peritoneal macrophages. Immunopharmacol Immunotoxicol 2012;34:303-308.
19. Franco-Molina MA, Mendoza-Gamboa E, Sierra-Rivera CA, Gómez-Flores RA, Zapata-Benavides P, Castillo-Tello P, et al. Antitumor activity of colloidal silver on MCF-7 human breast cancer cells. J Exp Clin Cancer Res 2010;29:148.
20. Poch E, Miñambres R, Mocholí E, Ivorra C, Pérez-Aragó A, Guerri C, et al. RhoE interferes with Rb inactivation and regulates the proliferation and survival of the U87 human glioblastoma cell line. Exp Cell Res 2007;313:719-731.
21. Senthilraja P, Kathiresan K. In vitro cytotoxicity MTT assay in Vero, HepG2 and MCF-7 cell lines study of Marine Yeast. J Appl Pharm Sci 2015;5:80-84.
22. Plumb JA (2004). Cell sensitivity assays: the MTT assay. Cancer Cell Culture. Humana Press, 2nd ed, Totowa, NJ, USA, pp. 165-169.
23. Araghi A, Hashemi S, Sepahi AA, Faramarzi MA, Amin M. Purification and study of anti-cancer effects of Serratia marcescens serralysin. Iran J Microbiol 2019;11:320-327.
24. Thon N, Thorsteinsdottir J, Eigenbrod S, Schüller U, Lutz J, Kreth S, et al. Outcome in unresectable glioblastoma: MGMT promoter methylation makes the difference. J Neurol 2017;264:350-358.
25. Scatena C, Roncella M, Di Paolo A, Aretini P, Menicagli M, Fanelli G, et al. Doxycycline, an inhibitor of mitochondrial biogenesis, effectively reduces cancer stem cells (CSCs) in early breast cancer patients: a clinical pilot study. Front Oncol 2018;8:452.
26. Lamb R, Fiorillo M, Chadwick A, Ozsvari B, Reeves KJ, Smith DL, et al. Doxycycline down-regulates DNA-PK and radiosensitizes tumor initiating cells: implications for more effective radiation therapy. Oncotarget 2015;6:14005-14025.
27. Jalali F, Sharifi M, Salehi R. Kefir induces apoptosis and inhibits cell proliferation in human acute erythroleukemia. Med Oncol 2016;33:7.
28. Rizk S, Maalouf K, Baydoun E. The antiproliferative effect of kefir cell-free fraction on HuT-102 malignant T lymphocytes. Clin Lymphoma Myeloma 2009;9 Suppl 3:S198-203.
29. Vidhyalakshmi R, Vallinachiyar C. Apoptosis of human breast cancer cells (MCF-7) induced by polysacccharides produced by bacteria. J Cancer Sci Ther 2013;5:31-34.
30. Abdulrahman SA, Abou El-E GA, Gamal HO. Overexpression and cytocidal activity of parasporin-1 from Bacillus thuringiensis against human cancer cell lines. J Microb Biochem Technol 2017;9:220-226.
31. St Jean AT, Swofford CA, Panteli JT, Brentzel ZJ, Forbes NS. Bacterial delivery of Staphylococcus aureus α-hemolysin causes regression and necrosis in murine tumors. Mol Ther 2014;22:1266-1274.
32. Kumar MV, Thippeswamy B, Raj PV. Cytotoxicity and anticancer studies of Bacillus cereus and Bacillus pumilus metabolites targeting human cancer cells. Appl Biochem Microbiol 2014;50:619-623.
33. Cao X, Wang AH, Jiao RZ, Wang CL, Mao DZ, Yan L, et al. Surfactin induces apoptosis and G (2)/M arrest in human breast cancer MCF-7 cells through cell cycle factor regulation. Cell Biochem Biophys 2009;55:163-171.
34. Aldeewan A, Zhang Y, Su L. Bacillus thuringiensis parasporins functions on cancer cells. Int J Pure Appl Biosci 2014;2:67-74.
| Files | ||
| Issue | Vol 13 No 3 (2021) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v13i3.6400 | |
| Keywords | ||
| Bacterial extract; Bacillus atrophaeus; Anticancer effect; Anti-microbial | ||
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How to Cite
1.
Asadi S, Soleimani N, Khosravi Babadi Z, Ebrahimipour GH. Isolation and identification of the bacterium producing antitumor and antimicrobial compounds derived from Iranian swamp frog (Rana ridibunda) skin. Iran J Microbiol. 2021;13(3):372-380.
