Investigating the anticancer effect of the lantibiotic, Lacticin 3147

Document Type : Research Paper

Authors

1 Ph.D Student of Microbiology,Department of Biology,Damghan Branch, Islamic Azad University, Damghan, Iran

2 Associate Professor of Biochemistry ,Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

3 Assistant Professor of Mycology, Department of Biology, Damghan Branch,Islamic Azad University,Damghan ,Iran.

4 Department of Biology, Parand Branch, Islamic Azad University, Parand, Iran.

Abstract

The most common type of cancer is skin cancer, especially in men. Lacticin 3147 is a bacteriocin from the lantibiotic family. Since it is similar to nisin, Lacticin was considered for its anticancer properties’ investigation. The aim was to investigate the effect of Lacticin 3147. The MTT Assay helped assess A431 cell survival and cytotoxicity following treatment with Lacticin 3147 after 24 and 48 hours. The cell apoptosis amounts were investigated by measuring the activity of caspases 8 and 9 in A431 cancer cells treated with the IC50 concentration of Lacticin 3147 after 24, 48, and 72 hours. The Boyden Chamber method evaluated the invasion ability of A431 cancer cells treated with a sublethal dose of Lacticin 3147. Specific primers were designed for proapoptotic genes bax, antiapoptotic bcl-2, and the reference gene -actin. The expression level in A431 cells treated with IC50 concentration of Lacticin 3147 was investigated using the quantitative Real-Time PCR method. Data were analyzed by SPSS 16 software and ANOVA statistical test (P<0.05).
Statistical analysis of the data obtained from the MTT test showed that the growth of cells treated with different concentrations of Lacticin 3147 decreased significantly (p<0.001). The inhibitory effect of Lacticin 3147 on the viability of A431 cells depends on the concentration and treatment time. As a result, a 0.3 µM concentration and 72-hour duration were the most consequential effects. Lacticin IC50 concentration was equal to 0.8 µM. The activity of caspases 8 and 9 in the treated A431 cancer cells increased in a time-dependent manner, and after 72 hours, it was 2.36 and 2.72 times compared to the control, respectively (p<0.001). The invasion activity of A431 cancer cells treated with the sublethal dose of Lacticin 3147 decreased in a dose-dependent manner. The lowest invasion activity was at concentration of 0.5 µM with minimal lethal effect (p<0.001).
The expression of the bax proapoptotic gene in A431 cells treated with IC50 concentration of Lacticin 3147 increased significantly (p<0.001). Also, the antiapoptotic bcl-2 gene expression reduction was significant in the same conditions. As a result, the use of Lacticin 3147 in the future may lead to a new strategy for the treatment of skin cancer.

Keywords


  1. Madan V, Lear JT, Szeimies R-M. Non-melanoma skin cancer. The Lancet. 2010;375(9715):673-85.https://doi.org/10.1016/S0140-6736(09)61196-X.
  2. Bray F, Laversanne M, Weiderpass E, Soerjomataram I. The ever‐increasing importance of cancer as a leading cause of premature death worldwide. Cancer. 2021.
  3. Narayanan DL, Saladi RN, Fox JL. Ultraviolet radiation and skin cancer. International journal of dermatology. 2010;49(9):978-86.
  4. Cotter PD, O'Connor PM, Draper LA, Lawton EM, Deegan LH, Hill C, et al. Posttranslational conversion of L-serines to D-alanines is vital for optimal production and activity of the lantibiotic lacticin 3147. Proceedings of the National Academy of Sciences. 2005;102(51):18584-9.
  5. Negash AW, Tsehai BA. Current applications of Bacteriocin. International Journal of Microbiology. 2020;2020.
  6. Frias MA, Somers S, Gerber-Wicht C, Opie LH, Lecour S, Lang U. The PGE2-Stat3 interaction in doxorubicin-induced myocardial apoptosis. Cardiovascular research. 2008;80(1):69-77.
  7. Chandran K, Aggarwal D, Migrino RQ, Joseph J, McAllister D, Konorev EA, et al. Doxorubicin inactivates myocardial cytochrome c oxidase in rats: cardioprotection by Mito-Q. Biophysical Journal. 2009;96(4):1388-98.
  8. Susin SA, Zamzami N, Kroemer G. Mitochondria as regulators of apoptosis: doubt no more. Biochimica et Biophysica Acta (BBA)-Bioenergetics. 1998;1366(1-2):151-65.
  9. Golstein P. Controlling cell death. Science. 1997;275(5303):1081-2.
  10. Freitas JT, Jozic I, Bedogni B. Wound healing assay for melanoma cell migration. Melanoma: Springer; 2021. p. 65-71.
  11. Kumar P, Nagarajan A, Uchil PD. Analysis of cell viability by the lactate dehydrogenase assay. Cold Spring Harbor Protocols. 2018;2018(6):pdb. prot095497.
  12. Forkasiewicz A, Dorociak M, Stach K, Szelachowski P, Tabola R, Augoff K. The usefulness of lactate dehydrogenase measurements in current oncological practice. Cellular & Molecular Biology Letters. 2020;25(1):1-14.
  13. Worsley CM, Veale RB, Mayne ES. Inducing apoptosis using chemical treatment and acidic pH, and detecting it using the Annexin V flow cytometric assay. Plos one. 2022;17(6):e0270599.
  14. Li D, Wu L-J, Tashiro S-i, Onodera S, Ikejima T. Oridonin inhibited the tyrosine kinase activity and induced apoptosis in human epidermoid carcinoma A431 cells. Biological and Pharmaceutical Bulletin. 2007;30(2):254-60.
  15. Montaser R, Luesch H. Marine natural products: a new wave of drugs? Future medicinal chemistry. 2011;3(12):1475-89.
  16. Imran M, Rauf A, Abu-Izneid T, Nadeem M, Shariati MA, Khan IA, et al. Luteolin, a flavonoid, as an anticancer agent: A review. Biomedicine & Pharmacotherapy. 2019;112:108612.https://doi.org/10.1016/j.biopha.2019.108612.
  17. Ryan A, Patel P, O'Connor PM, Ross RP, Hill C, Hudson SP. Pharmaceutical design of a delivery system for the bacteriocin lacticin 3147. Drug Deliv Transl Res. 2021;11(4):1735-51.10.1007/s13346-021-00984-9.
  18. Ochiiwa H, Ailiken G, Yokoyama M, Yamagata K, Nagano H, Yoshimura C, et al. TAS4464, a NEDD8-activating enzyme inhibitor, activates both intrinsic and extrinsic apoptotic pathways via c-Myc-mediated regulation in acute myeloid leukemia. Oncogene. 2021;40(7):1217-30.
  19. Zhang X, Bommareddy A, Chen W, Khalifa S, Kaushik RS, Fahmy H, et al. Sarcophine-diol, a Chemopreventive Agent of Skin Cancer, Inhibits Cell Growth and Induces Apoptosis through Extrinsic Pathway in Human Epidermoid Carcinoma A431 Cells. Transl Oncol. 2009;2(1):21-30.10.1593/tlo.08190.
  20. Chung J, Yoon Y-O, Lee JS, Ha TK, Ryu SM, Kim KH, et al. Inulin induces dendritic cells apoptosis through the caspase-dependent pathway and mitochondrial dysfunction. Biological and Pharmaceutical Bulletin. 2011;34(4):495-500.
  21. Juszczak AM, Czarnomysy R, Strawa JW, Zovko Končić M, Bielawski K, Tomczyk M. In vitro anticancer potential of Jasione montana and its main components against human amelanotic melanoma cells. International journal of molecular sciences. 2021;22(7):3345.
  22. Lin YS, Tsai PH, Kandaswami CC, Cheng CH, Ke FC, Lee PP, et al. Effects of dietary flavonoids, luteolin, and quercetin on the reversal of epithelial–mesenchymal transition in A431 epidermal cancer cells. Cancer science. 2011;102(10):1829-39.
  23. Muller S, Migianu E, Lecouvey M, Kraemer M, Oudar O. Alendronate inhibits proliferation and invasion of human epidermoid carcinoma cells in vitro. Anticancer research. 2005;25(4):2655-60.
  24. Sun W, Tao Y, Yu D, Zhao T, Wu L, Yu W, et al. Myricetin exerts potent anticancer effects on human skin tumor cells. Tropical Journal of Pharmaceutical Research. 2018;17(6):1067-72.
  25. Napoli S, Scuderi C, Gattuso G, Di Bella V, Candido S, Basile MS, et al. Functional roles of matrix metalloproteinases and their inhibitors in melanoma. Cells. 2020;9(5):1151.
  26. Bandyopadhyay D, Lopez G, Cantu S, Balboa S, Garcia A, Silva C, et al. Key Enzymes in Cancer: Mechanism of Action and Inhibition With Anticancer Agents. 2018.
  27. Webster MR, Fane ME, Alicea GM, Basu S, Kossenkov AV, Marino GE, et al. Paradoxical role for wild-type p53 in driving therapy resistance in melanoma. Molecular cell. 2020;77(3):633-44. e5.