Synergistic Anticancer Effects of Low-Frequency Magnetic Field and Doxorubicin on Glioblastoma Cell Line

Hilal Ergene; Murat Aydemir; Mehmet Enes Arslan; Gürkan Berber; Dilara Esra Men; Hatice Karataş; Elif Arslan; Cihat Aksakal; Hasan Türkez

Synergistic Anticancer Effects of Low-Frequency Magnetic Field and Doxorubicin on Glioblastoma Cell Line

Abstract

Glioblastoma (GBM) is one of the most aggressive primary brain tumors of the central nervous system, with a limited median survival of approximately 15 months despite current treatment options. In this study, the effects of a low-frequency magnetic field (LF-MF) in combination with the chemotherapeutic agent doxorubicin (DOX) on the glioblastoma cell line (U87MG) were investigated. In the experimental design, U87MG cells were exposed to LF-MF at intensity of 1 mT and treated with different concentrations of DOX (5 µg/ml and 10 µg/ml). Cell viability was assessed using the MTT assay, nuclear morphological changes were analyzed with Hoechst 33258 staining, and apoptotic and necrotic cell percentages were determined by flow cytometry. The results showed that DOX alone reduced cell viability in a dose-dependent manner (IC₅₀= 3.22 µg/ml). However, in the presence of LF-MF, DOX's cytotoxic effect increased, leading to a decrease in the IC₅₀ value to 2.18 µg/ml. Flow cytometry analyses revealed that while 5 µg/ml and 10 µg/ml DOX alone increased apoptotic cell percentages to 23.4% and 38.7%, respectively, these rates increased to 37.2% and 52.5% when combined with LF-MF. Compared to the control group, LF-MF did not alter toxicity in the healthy fibroblast cell line (HDFa) and had no additional effect on DOX-induced cell death. However, in glioblastoma cells, LF-MF-supported DOX treatment significantly enhanced cell death, suggesting that LF-MF may improve DOX efficacy by increasing cell membrane permeability and reactive oxygen species (ROS) production. This study demonstrates that the combination of LF-MF and DOX exerts a synergistic effect on glioblastoma cells by enhancing cell death. LF-MF-assisted chemotherapy strategies could be a potential alternative in glioblastoma treatment, but further molecular and biochemical studies are needed to elucidate the underlying mechanisms.

Keywords

Glioblastoma , Low-Frequency Magnetic Field , Doxorubicin , Apoptosis , Chemotherapy , Cell Viability

References

1. Türkiye Klinikleri. (2018). Biyofilm nedir? Türkiye Klinikleri. Erişim tarihi: 9 Aralık 2024, https://www.turkiyeklinikleri.com/article/en-what-is-biofilm--83796.html
2. Kartal, M., & Ekinci, M. (2021). Biyofilm yapısı ve önlenmesi. Akademik Gıda, Erişim tarihi: 9 Aralık 2024, https://dergipark.org.tr/en/pub/akademik-gida/article/1011231 3. Jefferson, K. K. (2004). What drives bacteria to produce a biofilm? FEMS Microbiology Letters, 236(2), 163–173. https://doi.org/10.1016/j.femsle.2004.06.005
4. Erkihun, M., Asmare, Z., Endalamew, K., Getie, B., Kiros, T., & Berhan, A. (2024). Medical scope of biofilm and quorum sensing during biofilm formation: Systematic review. Bacteria, 3(3), 118–135. https://doi.org/10.3390/bacteria3030008
5. Mirzaei, R., Abdi, M., & Gholami, H. (2020). The host metabolism following bacterial biofilm: What is the mechanism of action? Reviews and Research in Medical Microbiology, 31(4), 175–182. https://doi.org/10.1097/MRM.0000000000000216
6. Tarım ve Yaban Hayatı Bilimleri Dergisi. (2019). Güneydoğu Anadolu Bölgesinin farklı lokasyonlarından toplanan Boynuzlu Geven (Astragalus hamosus L.) otunun bazı kalite özelliklerinin belirlenmesi. Uluslararası Tarım ve Yaban Hayatı Bilimleri Dergisi, 5(2), 346–354. https://doi.org/10.24180/IJAWS.594960
7. Ekiz, G., Yılmaz, S., Yusufoglu, H., Kırmızıbayrak, P. B., & Bedir, E. (2019). Microbial transformation of cycloastragenol and astragenol by endophytic fungi isolated from Astragalus species. Journal of Natural Products, 82(11), 2979–2985. https://doi.org/10.1021/acs.jnatprod.9b00336
8. Li, X., et al. (2014). A review of recent research progress on the Astragalus genus. Molecules, 19(11), 18850–18880. https://doi.org/10.3390/molecules191118850
9. Abd Elkader, H. T. A. E., Essawy, A. E., & Al-Shami, A. S. (2022). Astragalus species: Phytochemistry, biological actions and molecular mechanisms underlying their potential neuroprotective effects on neurological diseases. Phytochemistry, 202, 113293. https://doi.org/10.1016/j.phytochem.2022.113293
10. Lavecchia, T., Rea, G., Antonacci, A., & Giardi, M. T. (2012). Healthy and adverse effects of plant-derived functional metabolites: The need of revealing their content and bioactivity in a complex food matrix. Critical Reviews in Food Science and Nutrition, 53(2), 198. https://doi.org/10.1080/10408398.2010.520829
11. Türkiye Florasında Yer Alan Endemik Astragalus Taksonları (Endemic Astragalus Taxa Found in Flora of Turkey). (2025, April 4). Erişim adresi: https://www.google.com/search?q=T%C3%BCrkiye+Floras%C4%B1nda+Yer+Alan+Endemik+Astragalus+Taksonlar%C4%B1+(...)

12. Li, Z., Qi, J., Guo, T., & Li, J. (2023). Research progress of Astragalus membranaceus in treating peritoneal metastatic cancer. Journal of Ethnopharmacology, 305, 116086. https://doi.org/10.1016/j.jep.2022.116086
13. Nafti, K., Giacinti, G., Marghali, S., & Raynaud, C. D. (2022). Screening for Astragalus hamosus triterpenoid saponins using HPTLC methods: Prior identification of azukisaponin isomers. Molecules, 27(17). https://doi.org/10.3390/molecules27175376
14. Li, X., et al. (2023). Regulation and mechanism of Astragalus polysaccharide on ameliorating aging in Drosophila melanogaster. International Journal of Biological Macromolecules, 234, 123632. https://doi.org/10.1016/j.ijbiomac.2023.123632
15. Piryaei, M., & Azimi, S. (2024). Preparation and evaluation of smart food packaging films with anthocyanin Sardasht black grape based on Astragalus gummifer and chitosan nanoparticles. International Journal of Biological Macromolecules, 254, 127974. https://doi.org/10.1016/j.ijbiomac.2023.127974
16. Risslegger, B., Lass-Flörl, C., Blum, G., & Lackner, M. (2015). Evaluation of a modified EUCAST fragmented-mycelium inoculum method for in vitro susceptibility testing of dermatophytes and the activity of novel antifungal agents. Antimicrobial Agents and Chemotherapy. https://doi.org/10.1128/AAC.04381-14
17. Haney, E. F., Trimble, M. J., Cheng, J. T., Vallé, Q., & Hancock, R. E. W. (2018). Critical assessment of methods to quantify biofilm growth and evaluate antibiofilm activity of host defence peptides. Biomolecules, 8(2), 29. https://doi.org/10.3390/biom8020029.
18. Makalesi, A., et al. (2021). Antimicrobial activity of pigments extracted from Auxenochlorella protothecoides SC3 against Pseudomonas aeruginosa. Tarım ve Doğa Dergisi, 10(2), 163–167. https://doi.org/10.46810/tdfd.930388
19. Schultz, A. W., Wang, J., Zhu, Z.-J., Johnson, C. H., Patti, G. J., & Siuzdak, G. (2013). Liquid chromatography quadrupole time-of-flight characterization of metabolites guided by the METLIN database. Nature Protocols, 8(3), 451–460. https://doi.org/10.1038/nprot.2013.004
20. Kumar, S. S., Manoj, P., & Giridhar, P. (2016). Fourier transform infrared spectroscopy (FTIR) analysis, chlorophyll content and antioxidant properties of native and defatted foliage of green leafy vegetables. Journal of Food Science and Technology, 53(4), 1874–1882. https://doi.org/10.1007/s13197-015-1959-0
21. Huang, W., et al. (2023). Integrating HPLC-Q-TOF-MS/MS, network pharmacology and experimental validation to decipher the chemical substances and mechanism of modified Gui-shao-liu-jun-zi decoction against gastric cancer. Journal of Traditional Chinese Medical Sciences, 13, 245–262. https://doi.org/10.1016/j.jtcme.2023.01.002 22. Bitki doku kültürlerinde sekonder metabolit sentezi. (Yıl yok). Gıda Dergisi. https://doi.org/10.15237/gida.GD13060
23. Salam, U., et al. (2023). Plant metabolomics: An overview of the role of primary and secondary metabolites against different environmental stress factors. Life, 13(3), 706. https://doi.org/10.3390/life13030706
24. Elshafie, H. S., Camele, I., & Mohamed, A. A. (2023). A comprehensive review on the biological, agricultural and pharmaceutical properties of secondary metabolites based-plant origin. International Journal of Molecular Sciences, 24(4), 3266. https://doi.org/10.3390/ijms24043266
25. Kandar, C. C. (2021). Secondary metabolites from plant sources. In Advanced Structured Materials (Vol. 140, pp. 329–377). https://doi.org/10.1007/978-3-030-54027-2_10 26. Bhatla, S. C., & Lal, M. A. (2023). Secondary metabolites. In Plant Physiology, Development and Metabolism (pp. 765–808). https://doi.org/10.1007/978-981-99-5736-1_33
27. Vaou, N., Stavropoulou, E., Voidarou, C., Tsigalou, C., & Bezirtzoglou, E. (2021). Microorganisms towards advances in medicinal plant antimicrobial activity: A review study on challenges and future perspectives. Microorganisms, 9(10), 2041.
28. Bocso, N., & Butnariu, M. (2022). The biological role of primary and secondary plants metabolites. Journal of Nutritional and Food Processing.
29. Zandavar, H., & Borhani, M. (2023). Secondary metabolites: Alkaloids and flavonoids in medicinal plants. IntechOpen.
30. Sharma, A., Sharma, S., Kumar, A., & Kaushik, V. K. (2022). Plant secondary metabolites: An introduction of their chemistry and biological significance with physicochemical aspect. In Springer Book Series.
31. Crozier, A., Clifford, M. N., & Ashihara, H. (2006). Plant secondary metabolites. Wiley. https://onlinelibrary.wiley.com/doi/abs/10.1002/9780470988558 32. Cui, L., Ma, Z., Li, W., Ma, H., Guo, S., Wang, D., & Niu, Y. (2023). Inhibitory activity of flavonoids fraction from Astragalus membranaceus Fisch. ex Bunge stems and leaves on Bacillus cereus and its separation and purification. Frontiers in Pharmacology, 14, 1183393.

Details

Primary Language

English

Subjects

Biochemistry and Cell Biology (Other)

Journal Section

Research Article

Authors

Hilal Ergene
0009-0008-1177-127X
Türkiye

Murat Aydemir
0000-0002-8359-5649
Türkiye

Mehmet Enes Arslan ^*
0000-0002-1600-2305
Türkiye

Gürkan Berber
0009-0006-4602-6443
Türkiye

Dilara Esra Men
0009-0009-5815-1673
Türkiye

Hatice Karataş
0009-0005-0920-8902
Türkiye

Elif Arslan
0000-0001-7310-241X
Türkiye

Cihat Aksakal
0009-0003-7519-5500
Türkiye

Hasan Türkez
0000-0002-7046-8990
Türkiye

Early Pub Date

July 21, 2025

Publication Date

July 13, 2025

Submission Date

March 25, 2025

Acceptance Date

May 11, 2025

Published in Issue

Year 2025 Volume: 4 Number: 1

IZ

https://izlik.org/JA77UZ69JK

APA

Ergene, H., Aydemir, M., Arslan, M. E., Berber, G., Men, D. E., Karataş, H., Arslan, E., Aksakal, C., & Türkez, H. (2025). Synergistic Anticancer Effects of Low-Frequency Magnetic Field and Doxorubicin on Glioblastoma Cell Line. Eurasian Journal of Molecular and Biochemical Sciences, 4(1), 25-36. https://izlik.org/JA77UZ69JK