Halotolerant Bacillus licheniformis Extract Induces Apoptosis in MCF-7 Cells via Modulation of BAX/BCL-2 Expression

Khoshandam, Mahdiyeh; Memarian, Mehrdad; Kalhor, Naser; Khoshandam, Mohadeseh; Soltaninejad, Hossein; Hedayati Goudarzi, Mohammad Taghi

year 19, Issue 6 (November - December 2025) Iran J Med Microbiol 2025, 19(6): 432-439 | Back to browse issues page

Mendeley

Zotero

RefWorks

Khoshandam M, Memarian M, Kalhor N, Khoshandam M, Soltaninejad H, Hedayati Goudarzi M T. Halotolerant Bacillus licheniformis Extract Induces Apoptosis in MCF-7 Cells via Modulation of BAX/BCL-2 Expression. Iran J Med Microbiol 2025; 19 (6) :432-439
URL: http://ijmm.ir/article-1-2778-en.html

Halotolerant Bacillus licheniformis Extract Induces Apoptosis in MCF-7 Cells via Modulation of BAX/BCL-2 Expression

Mahdiyeh Khoshandam¹

, Mehrdad Memarian²

, Naser Kalhor³

, Mohadeseh Khoshandam⁴

, Hossein Soltaninejad⁵

, Mohammad Taghi Hedayati Goudarzi⁶

1- Department of Genetics, Islamic Azad University, Qom, Iran
2- Department of Microbiology, Islamic Azad University, Qom Branch, Qom, Iran
3- Department of Cell Biology and Regenerative Medicine, Academic Center for Education, Culture and Research, Qom Branch, Qom, Iran
4- National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran
5- Department of Stem Cells Technology and Tissue Regeneration, Faculty of Interdisciplinary Science and Technologies, Tarbiat Modares University, Tehran, Iran , hosoltaninejad@gmail.com
6- Department of Cardiology, School of Medicine, Babol University of Medical Sciences, Babol, Iran

Abstract: (491 Views)

Background and Aim: Breast cancer is one of the leading causes of cancer-related deaths among women worldwide. The resistance of advanced tumors to conventional treatments highlights the need for alternative therapies. Apoptosis, regulated by key proteins like BAX (pro-apoptotic) and BCL-2 (anti-apoptotic), plays a crucial role in controlling cancer cell proliferation. This study aimed to investigate the effects of a halotolerant Bacillus (B.) licheniformis extract, isolated from Hoz-e Sultan Lake in Qom Province, on the expression of BAX and BCL-2 genes in MCF-7 breast cancer cell line.
Materials and Methods: MCF-7 cells were treated with various concentrations of the bacterial extract and cell viability was assessed using MTT assay. Staurosporine was used as positive control. RNA extraction, cDNA synthesis, and Real-Time PCR were performed to evaluate target genes expression levels. Statistical analysis was conducted using an independent t-test and P-values less than 0.05 were considered significant.
Results: The extract significantly increased BAX expression (P<0.05) at 24, 48, and 72 hr, while BCL-2 expression initially decreased but showed a slight increase at 48 hr. The IC₅₀ values decreased over time, indicating time-dependent cytotoxicity.
Conclusion: The halotolerant B. licheniformis extract induced apoptosis in MCF-7 cells through BAX/BCL-2 pathway modulation, suggesting its potential as an alternative therapeutic source. Further studies are needed to identify active compounds and validate efficacy in vivo.

Keywords: Apoptosis, Bacillus licheniformis, BAX, BCL-2, Breast Cancer, MCF-7

Full-Text [PDF 773 kb] (43 Downloads)

Type of Study: Original Research Article | Subject: Microbial Genetics
Received: 2025/10/15 | Accepted: 2025/12/12 | ePublished: 2025/12/29

References

1. Kim J, Harper A, McCormack V, Sung H, Houssami N, Morgan E, et al. Global patterns and trends in breast cancer incidence and mortality across 185 countries. Nat Med. 2025;31(4):1154-62. [DOI:10.1038/s41591-024-02887-x] [DOI:10.1038/s41591-025-03502-3] [PMID]

2. Khoshandam M, Soheili ZS, Hosseinkhani S, Samiee S, Latifi-Navid H, Ahmadieh H, et al. In vivo inhibition of angiogenesis by htsFLT01/MiRGD nano complex. Transl Oncol. 2025;56:102400. [DOI:10.1016/j.tranon.2025.102400] [PMID] [PMCID]

3. Farhangfar SD, Fesahat F, Zare-Zardini H, Dehghan-Manshadi M, Zare F, Miresmaeili SM, et al. In vivo study of anticancer activity of ginsenoside Rh2-containing arginine-reduced graphene in a mouse model of breast cancer. Iran J Basic Med Sci. 2022;25(12):1442.

4. Riaz N, Jeen T, Whelan TJ, Nielsen TO. Recent advances in optimizing radiation therapy decisions in early invasive breast cancer. Cancers. 2023;15(4):1260. [DOI:10.3390/cancers15041260] [PMID] [PMCID]

5. Obeagu EI, Obeagu GU. Breast cancer: A review of risk factors and diagnosis. Medicine. 2024;103(3):e36905. [DOI:10.1097/MD.0000000000036905] [PMID] [PMCID]

6. Yuan, J. and D. Ofengeim, A guide to cell death pathways. Nat Rev Mol Cell Biol. 2024;25(5): 379-95. [DOI:10.1038/s41580-023-00689-6] [PMID]

7. Fath-Bayati L, Naserpour L, Khoshandam M, Jannatifar R, Fazaeli H. Recent advances in developing 3D culture systems of spermatogonial stem cell preservation and differentiation: A narrative review. Int J Reprod BioMed. 2023;21(9):681. [DOI:10.18502/ijrm.v21i9.14397] [PMID] [PMCID]

8. Dandoti S. Mechanisms adopted by cancer cells to escape apoptosis-A review. Biocell. 2021;45(4):863. [DOI:10.32604/biocell.2021.013993]

9. Qian S, Wei Z, Yang W, Huang J, Yang Y, Wang J. The role of BCL-2 family proteins in regulating apoptosis and cancer therapy. Front Oncol. 2022;12:985363. [DOI:10.3389/fonc.2022.985363] [PMID] [PMCID]

10. Iksen, Witayateeraporn W, Hardianti B, Pongrakhananon V. Comprehensive review of Bcl‐2 family proteins in cancer apoptosis: Therapeutic strategies and promising updates of natural bioactive compounds and small molecules. Phytother Res. 2024;38(5):2249-75. [DOI:10.1002/ptr.8157] [PMID]

11. Bahadar N, Bahadar S, Sajid A, Wahid M, Ali G, Alghamdi A, et al. Epigallocatechin gallate and curcumin inhibit Bcl-2: a pharmacophore and docking based approach against cancer. Breast Cancer Res. 2024;26(1):114. [DOI:10.1186/s13058-024-01868-9] [PMID] [PMCID]

12. Zeigler DR, Perkins JB. The genus bacillus. InPractical handbook of microbiology. 2021 May 4 (pp. 249-278). CRC Press. [DOI:10.1201/9781003099277-24]

13. Lu S, Wang C, Ma J, Wang Y. Metabolic mediators: microbial-derived metabolites as key regulators of anti-tumor immunity, immunotherapy, and chemotherapy. Front Immunol. 2024;15:1456030. [DOI:10.3389/fimmu.2024.1456030] [PMID] [PMCID]

14. Giurini EF, Godla A, Gupta KH. Redefining bioactive small molecules from microbial metabolites as revolutionary anticancer agents. Cancer Gene Ther. 2024;31(2):187-206. [DOI:10.1038/s41417-023-00715-x] [PMID] [PMCID]

15. Shleeva MO, Kondratieva DA, Kaprelyants AS. Bacillus licheniformis: a producer of antimicrobial substances, including antimycobacterials, which are feasible for medical applications. Pharmaceutics. 2023;15(7):1893. [DOI:10.3390/pharmaceutics15071893] [PMID] [PMCID]

16. Ramirez-Olea H, Reyes-Ballesteros B, Chavez-Santoscoy RA. Potential application of the probiotic Bacillus licheniformis as an adjuvant in the treatment of diseases in humans and animals: A systematic review. Front Microbiol. 2022;13:993451. [DOI:10.3389/fmicb.2022.993451] [PMID] [PMCID]

17. Evelyn, Utami SP, Chairul. Effect of temperature and soluble solid on Bacillus subtilis and Bacillus licheniformis spore inactivation and quality degradation of pineapple juice. Food Sci Technol Int. 2022;28(4):285-96. [DOI:10.1177/10820132211019143] [PMID]

18. Espoui AH, Larimi SG, Darzi GN. Optimization of protease production process using bran waste using Bacillus licheniformis. Korean J Chem Eng. 2022;39(3):674-83. [DOI:10.1007/s11814-021-0965-3]

19. Li C, Liu Z, Zhao J, Zhao ZM. Mechanisms and strategies for regulating sporulation and germination of Bacillus subtilis to enhance probiotic effects. Ind Crops Prod. 2024;220:119178. [DOI:10.1016/j.indcrop.2024.119178]

20. Khodadoost M, Khoshandam M, Mirtorabi SD, Ghadirzadeh MR, Ahangari G. Gene Expression Survey of Serotonin Receptors (5HTR2A and 5HTR3A) and Monoamine Oxidases Enzyme After Treatment With Methadone in Addicted Individuals. Int J Med Toxicol Forensic Med. 2019;9(3):125-32. [DOI:10.32598/ijmtfm.v9i3.25480]

21. Khoshandam M, Soheili ZS, Hosseinkhani S, Samiee S, Latifi-Navid H, Kalhor N, et al. Leveraging the htsFLT01/MiRGD Complex to Enhance Apoptosis and Suppress Angiogenesis in MCF7 Breast Cancer Cells. Iran J Med Sci. 2025;50(10):707.

22. Mustafa M, Ahmad R, Tantry IQ, Ahmad W, Siddiqui S, Alam M, et al. Apoptosis: a comprehensive overview of signaling pathways, morphological changes, and physiological significance and therapeutic implications. Cells. 2024;13(22):1838. [DOI:10.3390/cells13221838] [PMID] [PMCID]

23. Saddam M, Paul SK, Habib MA, Fahim MA, Mimi A, Islam S, et al. Emerging biomarkers and potential therapeutics of the BCL-2 protein family: the apoptotic and anti-apoptotic context. Egypt J Med Hum Genet. 2024;25(1):12. [DOI:10.1186/s43042-024-00485-7]

24. Loftus LV, Amend SR, Pienta KJ. Interplay between cell death and cell proliferation reveals new strategies for cancer therapy. Int J Mol Sci. 2022;23(9):4723. [DOI:10.3390/ijms23094723] [PMID] [PMCID]

25. Khoshandam M, Naserpour L, Goudarzi MT, Soltaninejad H, Zare-Zardini H, Hoseiny AA, et al. Regulatory Roles of MicroRNAs in Female and Male Human Reproduction: A Narrative Review. Iran J Med Sci. 2025;50(9):588.

26. Jia LT, Chen SY, Yang AG. Cancer gene therapy targeting cellular apoptosis machinery. Cancer Treat Rev. 2012;38(7):868-76. [DOI:10.1016/j.ctrv.2012.06.008] [PMID]

27. Jabbari K, Khalafizadeh A, Sheikhbahaei M, Soltaninejad H, Babashah S. TET1: the epigenetic architect of clinical disease progression. Genes Dis. 2025;12(5):101513. [DOI:10.1016/j.gendis.2025.101513] [PMID] [PMCID]

28. Zaimy MA, Saffarzadeh N, Mohammadi A, Pourghadamyari H, Izadi P, Sarli A, et al. New methods in the diagnosis of cancer and gene therapy of cancer based on nanoparticles. Cancer Gene Ther. 2017;24(6):233-43. [DOI:10.1038/cgt.2017.16] [PMID]

29. Yang P, Khoshandam M, Bhia I, Raji S, Soltaninejad H, Hosseinkhani S, et al. Integrating CRISPR/Cas technology with clinical trials: principles, progress, and challenges. Asian J Pharm Sci. 2025:101068. [DOI:10.1016/j.ajps.2025.101068] [PMID] [PMCID]

30. Hwang KT, Kim YA, Kim J, Oh HJ, Park JH, Choi IS, et al. Prognostic influences of BCL1 and BCL2 expression on disease-free survival in breast cancer. Sci Rep. 2021;11(1):11942. [DOI:10.1038/s41598-021-90506-x] [PMID] [PMCID]

31. Placzek WJ, Wei J, Kitada S, Zhai D, Reed JC, Pellecchia M. A survey of the anti-apoptotic Bcl-2 subfamily expression in cancer types provides a platform to predict the efficacy of Bcl-2 antagonists in cancer therapy. Cell Death Dis. 2010;1(5):e40. [DOI:10.1038/cddis.2010.18] [PMID] [PMCID]

32. Markouli M, Pagoni MN, Diamantopoulos P. BCL-2 inhibitors in hematological malignancies: biomarkers that predict response and management strategies. Front Oncol. 2025;14:1501950. [DOI:10.3389/fonc.2024.1501950] [PMID] [PMCID]

33. Zhang L, Lu Z, Zhao X. Targeting Bcl-2 for cancer therapy. Biochim Biophys Acta Rev Cancer. 2021;1876(1):188569. [DOI:10.1016/j.bbcan.2021.188569] [PMID] [PMCID]

34. Chaudhry GE, Akim AM, Sung YY, Muhammad TS. Cancer and apoptosis. InApoptosis and cancer: methods and protocols 2022 Sep 11 (pp. 191-210). New York, NY: Springer US. [DOI:10.1007/978-1-0716-2553-8_16] [PMID]

35. Altonsy MO, Andrews SC, Tuohy KM. Differential induction of apoptosis in human colonic carcinoma cells (Caco-2) by Atopobium, and commensal, probiotic and enteropathogenic bacteria: mediation by the mitochondrial pathway. Int J Food Microbiol. 2010;137(2-3):190-203. [DOI:10.1016/j.ijfoodmicro.2009.11.015] [PMID]

36. Lobet E, Letesson JJ, Arnould T. Mitochondria: a target for bacteria. Biochem Pharmacol. 2015;94(3):173-85. [DOI:10.1016/j.bcp.2015.02.007] [PMID]

37. Kokoulin MS, Kuzmich AS, Filshtein AP, Prassolov VS, Romanenko LA. Capsular polysaccharide from the marine bacterium Cobetia marina induces apoptosis via both caspase-dependent and mitochondria-mediated pathways in HL-60 cells. Carbohydr Polym. 2025;347:122791. [DOI:10.1016/j.carbpol.2024.122791] [PMID]

38. Budu O, Mioc A, Soica C, Caruntu F, Milan A, Oprean C, et al. Lactiplantibacillus plantarum induces apoptosis in melanoma and breast cancer cells. Microorganisms. 2024;12(1):182. [DOI:10.3390/microorganisms12010182] [PMID] [PMCID]

39. Areewong S, Suppramote O, Prasopporn S, Jirawatnotai S. Exploiting acquired vulnerability to develop novel treatments for cholangiocarcinoma. Cancer Cell Int. 2024;24(1):362. [DOI:10.1186/s12935-024-03548-2] [PMID] [PMCID]