Effects of Long-Term Pulsed Electromagnetic Field Exposure on Apoptosis in Neuroblastoma Cell Line

Abstract

Pulsed Electromagnetic Field (PEMF) is a low-frequency electromagnetic field that has gained popularity in clinical research in recent years for its potential therapeutic benefits. Unlike studies evaluating the effects of PEMF exposure combined with chemotherapeutic drugs at different frequencies, intensities, wavelengths and durations on apoptosis in various cancer cells, including brain cancer cells, in the study, the possible effects of long-term (48 hours) PEMF exposure at constant frequency and intensity (50 Hz, 1 mT) on the apoptosis mechanism in human neuroblastoma cells have been investigated by different techniques. Cells group is divided into three experimental groups as control, SK-N-SH cell group without PEMF exposure, and SK-N-SH cell group exposed to PEMF for 48 hours. Alamar blue, flow cytometry, qRT-PCR and Western-Blotting techniques were utilized to measure cell viability, apoptosis, mRNA level and protein expression of the caspase-8. It has been shown that long-term PEMF exposure significantly reduces cell viability in human neuroblastoma cells, inducing the cells to a significant amount of early apoptosis, and this mechanism may be explained by the increased mRNA and protein expression level of the caspase-8.

Keywords

• Pulsed electromagnetic field
• Neuroblastoma
• Caspase-8

Nöroblastoma Hücre Hattında Uzun Süreli Darbeli Elektromanyetik Alan Maruziyetinin Apoptoz Üzerine Etkileri

Abstract

Darbeli Elektromanyetik Alan (PEMF) düşük frekanslı elektromanyetik alan olup son yıllarda klinik araştırmalarda tedavi amaçlı uygulanmasına yönelik çalışmalar hız kazanmıştır. Farklı frekans, yoğunluk, dalga boyu ve sürelerde kematerapötik ilaçlarla birlikte uygulanan PEMF maruziyetinin beyin kanseri hücreleri dahil çeşitli kanser hücrelerinde apoptoz üzerine etkilerini değerlendiren çalışmalardan farklı olarak çalışmada sabit frekans ve yoğunlukta (50 Hz, 1 mT) uzun süre (48 saat) PEMF maruziyetinin SK-N-SH insan nöroblastoma hücresinde apoptoz mekanizmasına olası etkileri farklı tekniklerle araştırılmıştır. Hücreler kontrol grubu, PEMF maruziyetinin olmadığı SK-N-SH hücre grubu, ve PEMF’ye 48 saat maruz bırakılan SK-N-SH hücre grubu olmak üzere üçe ayrılmıştır. Hücre canlılığı, apoptoz tayini, kaspaz-8 mRNA düzeyi ve kaspaz-8 protein ekspresyonu sırasıyla alamar mavisi, akış sitometri, qRT-PCR ve Western-Blot teknikleriyle belirlenmiştir. Uzun süreli PEMF maruziyetinin insan nöroblastoma hücresinde hücre canlılığını belirgin şekilde azaltıp hücreleri daha fazla erken apoptoza uğratarak hücreleri apoptoza sürüklediği ve bu mekanizmanın kaspaz-8 mRNA düzeyinde ve protein ekspresyon seviyesinde artışla ilişkili olabileceği gösterilmiştir.

Keywords

• Darbeli elektromanyetik alan
• Nöroblastoma
• Kaspaz-8

References

1. H. Wang and X. Zhang, “Magnetic fields and reactive oxygen species,” Int. J. Mol. Sci., 18(10), 2175- 2017. doi: 10.3390/ijms18102175.
2. T. Oda and T. Koike, “Magnetic field exposure saves rat cerebellar granule neurons from apoptosis in vitro,” Neurosci. Lett., 365(2), 83-86, 2004. doi: 10.1016/j.neulet.2004.04.068.
3. A. Krath, T. Klüter, M. Stukenberg, P. Zielhardt, H. Gollwitzer, N. Harrasser, J. Hausdorf, M. Ringeisen, and L. Gerdesmeyer, “Electromagnetic transduction therapy in non-specific low back pain: A prospective randomised controlled trial,” J. Orthop., 14(3), 410-415, 2017. doi: 10.1016/j.jor.2017.06.016.
4. M. Vadalà, A. Vallelunga, L. Palmieri, B. Palmieri, J. C. Morales-Medina, and T. Iannitti, “Mechanisms and therapeutic applications of electromagnetic therapy in Parkinson's disease,” Behav. Brain Funct., 11, 26, 2015. doi: 10.1186/s12993-015-0070-z.
5. E. Capelli, F. Torrisi, L. Venturini, M. Granato, L. Fassina, G. F. D. Lupo, and G. Ricevuti, “Low-frequency pulsed electromagnetic field is able to modulate miRNAs in an experimental cell model of Alzheimer’s disease,” Journal of Healthcare Engineering, (5-6), 1-10, 2017.
6. M. Markov, “XXIst century magnetotherapy,” Electromagn. Biol. Med., 34(3), 190-196, 2015.
7. M. O. Mattsson, and M. Simkó, “Emerging medical applications based on non-ionizing electromagnetic fields from 0 Hz to 10 THz,” Medical Devices: Evidence and Research, 12, 347-368, 2019.
8. S. Gessi, S. Merighi, S. Bencivenni, E. Battistello, F. Vincenzi, S. Setti, M. Cadossi, P. A. Borea, R. Cadossi, and K. Varani, “Pulsed electromagnetic field and relief of hypoxia‐induced neuronal cell death: The signaling pathway,” J. Cell Physiol., 234, 15089-15097, 2019.

9. C. Daish, R. Blanchard, K. Fox, P. Pivonka, and E. Pirogova, “The application of pulsed electromagnetic fields (PEMFs) for bone fracture repair: past and perspective findings,” Annals of Biomedical Engineering, 46(4), 525-542, 2018.
10. B. Strauch, C. Herman, R. Dabb, L. J. Ignarro, and A. A. Pilla, “Evidence-based use of pulsed electromagnetic field therapy in clinical plastic surgery,” Surgery Journal, 29(2), 135 143, 2009.
11. T. Şimşek, S. Karakurt, and Ç. Gökçek Saraç, “Yüksek doz hidrojen peroksit ile muamele edilen insan nöroblastoma hücre hattında darbeli elektromanyetik alan maruziyetinin glutatyon miktarına etkisi,” Süleyman Demirel Üniversitesi Fen Edebiyat Fakültesi Fen Dergisi, 17(1), 146-154, 2022. doi: 10.29233/sdufeffd.1029835.
12. F. Vincenzi, A. Ravani, S. Pasquini, M. Merighi, S. Gessi, S. Setti, R. Cadossi, P. A. Borea, and K. Varani, “Pulsed electromagnetic field exposure reduces hypoxia and inflammation damage in neuron-like and microglial cells,” Journal Cellular Physiogy, 232, 1200-1208, 2017.
13. S. Falone, N. Marchesi, C. Osera, L. Fassina, S. Comincini, M. Amadio, and A. Pascale, “Pulsed electromagnetic field (PEMF) prevents pro-oxidant effects of H2O2 in SK-N-BE (2) human neuroblastoma cells,” International Journal of Radiation Biology, 92(5), 281-286, 2016.
14. S. Goldar, M. S. Khaniani, S. M. Derakhshan, and B. Baradaran, “Molecular mechanisms of apoptosis and roles in cancer development and treatment,” Asian Pac. J. Cancer Prev., 16(6), 2129-2144, 2015. doi: 10.7314/apjcp.2015.16.6.2129.
15. G. Atagün, Z. Eren, and İ. Gürkanlı, “Apoptoziste mitokondrinin rolü”, Türk Blimsel Derlemeler Dergisi, 4(2), 49-53, 2011.
16. G. Pistritto, D. Trisciuoglio, C. Ceci, A. Garufi, and G. D'Orazi, “Apoptosis as anticancer mechanism: function and dysfunction of its modulators and targeted therapeutic strategies,” Aging (Albany NY), 8(4), 603-619, 2016. doi: 10.18632/aging.100934.
17. A. W. Roberts, J. F. Seymour, J. R. Brown, W. G. Wierda, T. J. Kipps, S. L. Khaw, D. A. Carney, S. Z. He, D. C. S. Huang, H. Xiong, Y. Cui, T. A. Busman, E. M. McKeegan, A. P. Krivoshik, S. H. Enschede, and R. Humerickhouse, “Substantial susceptibility of chronic lymphocytic leukemia to BCL2 inhibition: results of a phase I study of navitoclax in patients with relapsed or refractory disease,” Journal of Clinical Oncology, 30(5), 488-496, 2011. doi: 10.1200/JCO.2011.34.7898.2011.
18. Z. Akbarnejad, H. Eskandary, L. Dini, C. Vergallo, S. N. Nematollahi-Mahani, A. Farsinejad, M. F. S. Abadi, and M. Ahmadi, “Cytotoxicity of temozolomide on human glioblastoma cells is enhanced by the concomitant exposure to an extremely low-frequency electromagnetic field (100Hz, 100G),” Biomed. Pharmacother., 92, 254-264, 2017. doi: 10.1016/j.biopha.2017.05.050.
19. M. Amiri, , M. Basiri, H. Eskandary, Z. Akbarnejad, M. Esmaeeli, Y. M. Ardakani, and M. Ahmadi-Zeidabadi, ‘ʻCytotoxicity of carboplatin on human glioblastoma cells is reduced by the concomitant exposure to an extremely low-frequency electromagnetic field (50 Hz, 70 G),” Electromagnetic Biology and Medicine, 37(3), 138-145, 2018. doi: 10.1080/15368378.2018.1477052.
20. T. Mercantepe, L. Tümkaya, M. F. Gökçe, Z. S. Topal, and E. Esmer, “900 MHz elektromanyetik alanın serebellum üzerine etkilerinin histopatolojik olarak incelenmesi,” Med. Bull. Sisli Etfal Hosp., 52(2), 129-134, 2018.
21. M. Z. Tuysuz, H. Kayhan, A. S. Yar-Saglam, E. U. Bagriacik, M. Yagci, and A. G. Canseven, “Cep telefonu maruziyetinden kaynaklanan radyofrekans elektromanyetik alanın apoptoz üzerine etkisi,” Harran Üniversitesi Tıp Fakültesi Dergisi, 16(1),123-129, 2019.
22. M. Barati, H. Fahimi, L. Farahmand, and A. Madjid-Ansari, “1Hz 100mT electromagnetic field induces apoptosis in breast cancer cells through up-regulation of P38 and P21,” Multidisciplinary Cancer Investigation, 4(1), 23-29, 2020. doi: 10.30699/acadpub.mci.4.1.23.
23. A. Xu, Q. Wang, and T. Lin, “Low-frequency magnetic fields (LF-MFs) inhibit proliferation by triggering apoptosis and altering cell cycle distribution in breast cancer cells”, Int. J. Mol. Sci., 21(8), 2952, 2020. doi: 10.3390/ijms21082952.
24. J. Kaszuba-Zwoinska, K. Wojcik, M. Bereta, A. Ziomber, P. Pierzchalski, E. Rokita, J. Marcinkiewicz, W. Zaraska, and P. Thor, “Pulsating electromagnetic field stimulation prevents cell death of puromycin treated U937 cell line, ” Journal of Physiology and Pharmacology, 61(2), 201-205, 2010.
25. F. Pasi, L. Fassina, M.E. Mognaschi, G. Lupo, F. Corbella, R. Nano, and E. Capelli, “Pulsed electromagnetic field with temozolomide can elicit an epigenetic pro-apoptotic effect on glioblastoma T98G cells,” Anti Cancer Research, 36, 5821-5826, 2016. doi:10.21873/anticanres.11166.
26. M. Jiang, L. Qi, L. Lisha, Y. Wu, D. Song, and Y. Yanjing, “Caspase-8: A key protein of cross-talk signal way in “PANoptosis” in cancer,” Int. J. Cancer, 149, 1408-1420, 2021.
27. M. Fritsch, S.D. Günther, R. Schwarzer, M.C. Albert, F. Schorn, J.P. Werthenbach, L.M. Schiffmann, N. Stair, H. Stocks, J.M. Seeger, M. Lamkanfi, M. Krönke, M. Pasparakis, and H. Kashkar, “Caspase-8 is the molecular switch for apoptosis, necroptosis and pyroptosis,” Nature, 575(7784), 683-687, 2019. doi: 10.1038/s41586-019-1770-6.
28. D.R. McIlwain, T. Berger, and T.W. Mak, “Caspase functions in cell death anddisease,” Cold Spring Harb Perspect Biol. 7(4), a026716, 2015. A. Bildik and İ. Bayar, “Kanserde apoptotik yolakların inhibisyonu,” Turkiye Klinikleri J Vet Sci. 9(2):42-51, 2018. doi: 10.5336/vetsci.2018-62141.