The effect of Extrmely low frequency magnetic field with Avastin on proliferation of Human umbilical vein endothelial cells

Zeraati, Vahid; Abdolmaleki, Parviz; Hassan Sajedi, Reza; Moazeni-roodi, Abdolkarim

The effect of Extrmely low frequency magnetic field with Avastin on proliferation of Human umbilical vein endothelial cells

Document Type : Original Research

Authors

vahid zeraati ¹

Parviz Abdolmaleki ¹

Reza Hassan Sajedi ²

Abdolkarim Moazeni-roodi ³

¹ Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University (TMU), Tehran, Iran

² Department of Biochemistry, Faculty of Biological Sciences , Tarbiat Modares University , Tehran , Iran

³ Department of Clinical Biochemistry,School of Medicine, Iranshahr University of Medical Sciences, Iranshahr, Iran

Abstract

Investigation of factors affecting endothelial cell proliferation is an essential part of angiogenesis studies. Given the importance of inhibiting angiogenesis in the treatment of cancers, and due to the side effects and high cost of anti-angiogenic drugs such as Avastin, the use of physical agents to aid in treatment and reduce the need for high doses of the drug is noteworthy. Magnetic fields are of interest due to their long-distance and non-invasive effects, and many studies have been conducted on their effects on biological phenomena, including angiogenesis, with inconsistent results. In the present study, the effect of a 2 mT alternating magnetic field with a frequency of 200 Hz and Austin on the proliferation of human umbilical vein endothelial cells (HUVEC) was investigated. Cells were treated for 48 hours under a mixture of 50 μg/ml solution of vascular endothelial growth factor (VEGEF) and Avastin at concentrations (zero (drug control), 50, 100, 200 and 400 μg/ml) as well as field treatment groups for They were exposed to magnetic fields for 3, 6, 12, 24 and 48 hours. Then, cell proliferation was assessed using Alamar Blue colorimetric test. Data were analyzed by three-way analysis of variance. According to the findings, the exposure times of 12, 24 and 48 hours showed a significant reduction in cell proliferation compared to the control group, but this difference was not significant in the 3 and 6 hour treatments. Also, the degree of interaction of these factors with each other on HUVEC proliferation was investigated.

Keywords

Magnetic Fields

Angiogenesis

Avastin

Vascular Endothelial Growth Factor

Human Umbilical Vein Endithelial Cells

20.1001.1.23222115.1400.13.1.6.5

Subjects

Industrial Biotechnology

1. Folkman, J., Tumor angiogenesis: therapeutic implications. N Engl J Med, 1971. 285(21): p. 1182-6.
2. Folkman, J., Role of angiogenesis in tumor growth and metastasis. Semin Oncol, 2002. 29(6 Suppl 16): p. 15-8.
3. Folkman, J. and Y. Shing, Angiogenesis. J Biol Chem, 1992. 267(16): p. 10931-4.
4. Zucchelli, E., Q.A. Majid, and G. Foldes, New artery of knowledge: 3D models of angiogenesis. Vascular Biology, 2019. 1(1): p. H135-H143.
5. Cines, D.B., et al., Endothelial Cells in Physiology and in the Pathophysiology of Vascular Disorders. Blood, 1998. 91(10): p. 3527-3561.
6. Yoo, S.Y. and S.M. Kwon, Angiogenesis and Its Therapeutic Opportunities. Mediators of Inflammation, 2013. 2013: p. 127170.
7. Al-Husein, B., et al., Antiangiogenic therapy for cancer: an update. Pharmacotherapy, 2012. 32(12): p. 1095-1111.
8. Zhang, W., et al., The Benefits and Side Effects of Bevacizumab for the Treatment of Recurrent Ovarian Cancer. Current drug targets, 2016. 18.
9. Rovithi, M. and H.M.W. Verheul, Pulsatile high-dose treatment with antiangiogenic tyrosine kinase inhibitors improves clinical antitumor activity. Angiogenesis, 2017. 20(3): p. 287-289.
10. Sengupta, S. and V.K. Balla, A review on the use of magnetic fields and ultrasound for non-invasive cancer treatment. Journal of Advanced Research, 2018. 14: p. 97-111.
11. de Seze, R., et al., Effects of 100 mT time varying magnetic fields on the growth of tumors in mice. Bioelectromagnetics, 2000. 21(2): p. 107-111.
12. Markov, M.S., Pulsed electromagnetic field therapy history, state of the art and future. The Environmentalist, 2007. 27(4): p. 465-475.
13. Delle Monache, S., et al., Inhibition of angiogenesis mediated by extremely low-frequency magnetic fields (ELF-MFs). PLoS One, 2013. 8(11): p. e79309.
14. Mostafaie, A., H.R.M. Motlagh, and K. Mansouri, Angiogenesis and the Models to Study Angiogenesis. Yakhteh Medical Journal, 2010. 11(4).
15. Folkman, J., Angiogenesis research: from laboratory to clinic. Forum (Genova), 1999. 9(3 Suppl 3): p. 59-62.
16. Balyasnikova, I., K. Krotov, and S. Danilov, Effect of a static magnetic field on the growth rate and in vitro angiogenesis of endothelial cells. Bulletin of experimental biology and medicine, 1994. 117(1): p. 110-113.
17. Potenza, L., et al., Effects of a 300 mT static magnetic field on human umbilical vein endothelial cells. Bioelectromagnetics, 2010. 31(8): p. 630-639.
18. Fei, L., et al., Effects of static magnetic field on human umbilical vessel endothelial cell. Journal of Medical Colleges of PLA, 2007. 22(2): p. 106-110.
19. Wang, Z., et al., Inhibitory effects of a gradient static magnetic field on normal angiogenesis. Bioelectromagnetics: Journal of the Bioelectromagnetics Society, The Society for Physical Regulation in Biology and Medicine, The European Bioelectromagnetics Association, 2009. 30(6): p. 446-453.
20. Martino, C.F., et al., Effects of weak static magnetic fields on endothelial cells. Bioelectromagnetics: Journal of the Bioelectromagnetics Society, The Society for Physical Regulation in Biology and Medicine, The European Bioelectromagnetics Association, 2010. 31(4): p. 296-301.
21. Martino, C.F., Static magnetic field sensitivity of endothelial cells. Bioelectromagnetics, 2011. 32(6): p. 506-508.
22. Yen‐Patton, G.A., et al., Endothelial cell response to pulsed electromagnetic fields: stimulation of growth rate and angiogenesis in vitro. Journal of cellular physiology, 1988. 134(1): p. 37-46.
23. Li, F., et al., Pulsed magnetic field accelerate proliferation and migration of cardiac microvascular endothelial cells. Bioelectromagnetics, 2015. 36(1): p. 1-9.
24. Hopper, R.A., et al., Osteoblasts stimulated with pulsed electromagnetic fields increase HUVEC proliferation via a VEGF-A independent mechanism. Bioelectromagnetics, 2009. 30(3): p. 189-197.
25. Delle Monache, S., et al., Extremely low frequency electromagnetic fields (ELF‐EMFs) induce in vitro angiogenesis process in human endothelial cells. Bioelectromagnetics: Journal of the Bioelectromagnetics Society, The Society for Physical Regulation in Biology and Medicine, The European Bioelectromagnetics Association, 2008. 29(8): p. 640-648.
26. Naarala, J., et al., Direction-dependent effects of combined static and ELF magnetic fields on cell proliferation and superoxide radical production. BioMed research international, 2017. 2017.
27. Wachsberger, P., R. Burd, and A.P. Dicker, Tumor response to ionizing radiation combined with antiangiogenesis or vascular targeting agents: exploring mechanisms of interaction. Clinical cancer research, 2003. 9(6): p. 1957-1971.
28. Dings, R.P.M., et al., Scheduling of radiation with angiogenesis inhibitors anginex and Avastin improves therapeutic outcome via vessel normalization. Clinical cancer research : an official journal of the American Association for Cancer Research, 2007. 13(11): p. 3395-3402.
29. Nieder, C., et al., Radiation therapy plus angiogenesis inhibition with bevacizumab: rationale and initial experience. Reviews on recent clinical trials, 2007. 2(3): p. 163-168.
30. Peng, F., et al., Recombinant human endostatin normalizes tumor vasculature and enhances radiation response in xenografted human nasopharyngeal carcinoma models. PloS one, 2012. 7(4): p. e34646.
31. Hsu, H.-W., et al., Combination antiangiogenic therapy and radiation in head and neck cancers. Oral Oncology, 2014. 50(1): p. 19-26.
32. Zhuang, H., et al., A study on the evaluation method and recent clinical efficacy of bevacizumab on the treatment of radiation cerebral necrosis. Scientific reports, 2016. 6: p. 24364-24364.
33. Gu, S., et al., Evaluating the effect of Avastin on breast cancer angiogenesis using synchrotron radiation. Quantitative imaging in medicine and surgery, 2019. 9(3): p. 418-426.
34. مشتاق, ص., et al., اثر عصاره آبی زعفران و میدان الکترو مغناطیس با فرکانس کم بر آنژیوژنز در حلقه آئورت موش صحرایی نژاد ویستار. کومش, 1393. 15(4 (پیاپی 52)): p. -.
35. Zafar-Balanezhad, S., et al., The synergic effects of rapamycin and extremely low frequency electromagnetic field on angiogenesis. Journal of Shahrekord Uuniversity of Medical Sciences, 2009. 11(3): p. 70-76.
36. مشتاق, et al., کاربرد توام پروتئازگیاهی بروملین و میدان الکترومغناطیس با فرکانس پایین ((50 HZ بر آنژیوژنز در مدل حلقه آئورت موش صحرایی. فیزیولوژی و تکوین جانوری, 2018. 12(شماره 1 زمستان 1397): p. 13-24.
37. Absher, M., CHAPTER 1 - Hemocytometer Counting, in Tissue Culture, P.F. Kruse and M.K. Patterson, Editors. 1973, Academic Press. p. 395-397.
38. Green, M.R. and J. Sambrook, Estimation of cell number by hemocytometry counting. Cold Spring Harbor Protocols, 2019. 2019(11): p. pdb. prot097980.
39. Koyanagi, M., S. Kawakabe, and Y. Arimura, A comparative study of colorimetric cell proliferation assays in immune cells. Cytotechnology, 2016. 68(4): p. 1489-1498.
40. Riss, T.L., et al., Cell viability assays, in Assay Guidance Manual [Internet]. 2016, Eli Lilly & Company and the National Center for Advancing Translational Sciences.
41. Al-Nasiry, S., et al., The use of Alamar Blue assay for quantitative analysis of viability, migration and invasion of choriocarcinoma cells. Human Reproduction, 2007. 22(5): p. 1304-1309.
42. Carneiro, A., et al., Multiple effects of bevacizumab in angiogenesis: implications for its use in age‐related macular degeneration. Acta ophthalmologica, 2009. 87(5): p. 517-523.
43. Wang, Y., et al., Biological activity of bevacizumab, a humanized anti-VEGF antibody in vitro. Angiogenesis, 2004. 7(4): p. 335-345.
44. Notara, M., et al., Bevacizumab Induces Upregulation of Keratin 3 and VEGFA in Human Limbal Epithelial Cells in Vitro. Journal of Clinical Medicine, 2019. 8(11): p. 1925.
45. Kleijn, A., et al., A Systematic Comparison Identifies an ATP-Based Viability Assay as Most Suitable Read-Out for Drug Screening in Glioma Stem-Like Cells. Stem Cells International, 2016. 2016: p. 5623235.
46. Li, F., et al., Effects of static magnetic field on human umbilical vessel endothelial cell. Journal of Medical Colleges of PLA, 2007. 22(2): p. 106-110.
47. Mahna, A. and M. Firoozabadi, Environmental 50Hz Magnetic Fields Can Increase Viability of Human Umbilical Vein Endothelial Cells (HUVEC). Iranian Journal of Medical Physics, 2016. 13.
48. Naarala, J., et al., Direction-Dependent Effects of Combined Static and ELF Magnetic Fields on Cell Proliferation and Superoxide Radical Production. BioMed research international, 2017. 2017: p. 5675086-5675086.
49. Madkan, A., et al., Steps to the clinic with ELF EMF. Natural Science, 2009. Vol.01No.03: p. 9.
50. Blank, M. and R. Goodman, Electromagnetic fields stress living cells. Pathophysiology, 2009. 16(2-3): p. 71-78.
51. Funk, R.H.W., T. Monsees, and N. Özkucur, Electromagnetic effects – From cell biology to medicine. Progress in Histochemistry and Cytochemistry, 2009. 43(4): p. 177-264.
52. Salari, V., et al., Towards non-invasive cancer diagnostics and treatment based on electromagnetic fields, optomechanics and microtubules. arXiv preprint arXiv:1708.08339, 2017.
53. Yuan, J., F. Xin, and W. Jiang, Underlying Signaling Pathways and Therapeutic Applications of Pulsed Electromagnetic Fields in Bone Repair. Cellular Physiology and Biochemistry, 2018. 46(4): p. 1581-1594.
54. Furse, C., D.A. Christensen, and C.H. Durney, Basic introduction to bioelectromagnetics. Third ed. 2018: CRC press.
55. Pall, M.L., Electromagnetic fields act via activation of voltage-gated calcium channels to produce beneficial or adverse effects. Journal of cellular and molecular medicine, 2013. 17(8): p. 958-965.
56. Wood, A. and K. Karipidis, Radiofrequency Fields and Calcium Movements Into and Out of Cells. Radiation Research, 2020. 195(1): p. 101-113.
57. Stovbun, S.V., et al., The weak magnetic field inhibits the supramolecular self-ordering of chiral molecules. Scientific Reports, 2020. 10(1): p. 17072.
58. Markov, M.S., Expanding Use of Pulsed Electromagnetic Field Therapies. Electromagnetic Biology and Medicine, 2007. 26(3): p. 257-274.
59. Lai, H., Exposure to Static and Extremely-Low Frequency Electromagnetic Fields and Cellular Free Radicals. Electromagnetic Biology and Medicine, 2019. 38(4): p. 231-248.
60. Mattei, M.D., et al., Effects of Pulsed Electromagnetic Fields on Human Articular Chondrocyte Proliferation. Connective Tissue Research, 2001. 42(4): p. 269-279.