Treatment of Cancer by Radiotherapy and Nanoparticles Coupled With Methotrexate

Document Type : Original Paper


1 Physics Department, Faculty of Science, Damanhour University, Egypt.

2 Physics Department, Faculty of Applied Medical Science, Pharos University, Egypt.

3 Physics Department, Faculty of Science, Damanhur University, Egypt


Introduction: Cancer patients receive radiation therapy (RT) as part of their treatment as monotherapy or as part of a combination treatment. Radiation therapy uses high-energy radiation such as photons and strong ions. Nanoparticles (NPs) are used for magnetic hyperthermia, which increases the efficacy of RT and generates heat to kill cancer cells by destroying their DNA.           
Material and Methods: Nanoparticles were prepared using the co-precipitation method and characterized using transmission electron microscopy (TEM), scanning electron microscopy (SEM), and X-ray diffractometer. Fifty-six male mice were housed under similar environmental conditions. The animals were injected into the right flank with 0.25 mL of 106 cells/mL Ehrlich tumor suspension. When tumors reached 5-10 mm in diameter, the mice were randomly divided into eight groups as follows: 1st group: used as the control group injected with 25 μL of phosphate buffer saline without treatment, 2nd group: injected with Fe3O4-NPs, 3rd group: injected with Fe2O3-NPs, 4th group: injected with methotrexate (MTX), 5thgroup: injected with MTX, Fe3O4, and Fe2O3-NPs, 6th group: as the 5th group with microwave hyperthermia, 7th group: as the 5th group then treated with high-energy photons, and 8th group: as the 5th group and exposed to electron beam therapy. Tumor volume and weight were measured after 15 days. Tumor apoptosis was studied using histopathology, and the tumor's side effects on the biological systems were investigated.
Results: The results indicated that magnetic hyperthermia with microwave and linear accelerator treatment coupled with drugs was suitable for cancer treatment. A significant decrease in tumor size, tumor necrosis, and fibrosis was observed.
Conclusion: It was found that Fe3O4 and Fe2O3-NPs coupled with MTX and exposure to photon and electron beam therapy and microwave radiation are the best methods for cancer treatment.   


Main Subjects


    1. Piumi Y, Liyanage D, Hettiarachchi Z, Allal Ouhtit, Elif S, Cagri O, et al. Nanoparticle-mediated targeted drug delivery for breast cancer treatment. Biochim Biophys Acta Rev Cancer. 2019 April; 1871(2): 419–33.
    2. Wust P, Hildebrandt B, Sreenivasa G, Rau B, Gellermann J, Riess H, et al. Hyperthermia in combined treatment of cancer. Lancet Oncol. 2002; 3: 487–7.
    3. Laurent S, Forge D, Port M. Magnetic iron oxide nanoparticles: synthesis, stabilization, vectorization, physic chemical characterizations and biological applications. Chemical Reviews. 2008;108(6): 2064–10.
    4. Laurent S, Dutz S, Hafeli UO, Mahmoudi M. Magnetic fluid hyperthermia: focus on superparamagnetic iron oxide nanoparticles. Advances in Colloid and Interface Science. 2011; 166(1-2): 8-23.
    5. Nosrati H. Anticancer activity of tamoxifen loaded tyrosine decorated biocompatible Fe3O4 magnetic nanoparticles against breast cancer cell lines. J Inorg Organomet Polym Mater. 2018;28(3):1178–86.
    6. Sanjay K, Pratibha K, Rajeev S Emerging. Nanomaterials for Cancer Therapy. Nanoparticles in Medicine. 2020: 25-54.
    7. Cunningham CH, Arai T, Yang PC, McConnell MV, Pauly JM, Conolly SM. Positive contrast magnetic resonance imaging of cells labeled with magnetic nanoparticles. Magnetic Resonance in Medicine. 2005;53(5): 999–05.
    8. Anderson SA, Rader R K, Westlin WF. Magnetic resonance contrast enhancement of neovasculature with alpha(v)beta(3)-targeted nanoparticles. Magnetic Resonance in Medicine. 2000;44 (3): 433–9.
    9. Polyak B, Friedman G. Magnetic targeting for site-specific drug delivery: applications and clinical potential. Expert Opinion on Drug Delivery. 2009 ;6(1): 53-70.
    10.  Weissleder R, Cheng HC, Bogdanova A, Bogdanov A. Magnetically labeled cells can be detected by MR imaging. Journal of Magnetic Resonance Imaging. 1997;7(1):258-63.
    11. Schellenberger EA, Reynolds F, Weissleder R, Josephson L. Surface-functionalized nanoparticle library yields probes for apoptotic cells. ChemBioChem. 2004;5(3):275–79.
    12.  Jalilian AR, Panahifar A, Mahmoudi M, Akhlaghi M, Simchi A. Preparation and biological evaluation of [67Ga]-labeled- superparamagnetic nanoparticles in normal rats. Radiochimica Acta. 2009 ;97(1): 51-6.
    13.  Corot C, Robert P, Idee JM, Port M. Recent advances in iron oxide nanocrystal technology for medical imaging. Advanced Drug Delivery Reviews. 2006;58(14): 1471-04.
    14.  Fan C, Gao W, Chen Z. Tumor selectivity of stealth multi-functionalized superparamagnetic iron oxide nanoparticles. International Journal of Pharmaceutics. 2011; 404(1-2): 180–90.
    15.  Jin G, He R, Liu Q, Dong Y, Lin M, Li W, Xu F. Theranostics of triple-negative breast cancer based on conjugated polymer nanoparticles. ACS Appl. Mater. Interfaces2018;10 :10634–46.
    16.  Park K, Lee S, Kang E, Kim K, Choi K, Kwon IC. New generation of multifunctional nanoparticles for cancer imaging and therapy. Adv Funct Mater. 2009;19(10):1553-66.
    17.  Atefeh R, Ameneh S.Gold nanoparticles as cancer theranostic agents. Nanomed J. 2019; 6(3): 147-60.
    18.  Pankhurst QA, Connolly J, Jones SK, Dobson J. Applications of magnetic nanoparticles in biomedicine. Journal of Physics D.2003;36(13):167–81.
    19.  Gilchrist R K, Shorey WD, Hanselman RC, Parrott JC, Taylor CB. Selective inductive heating of lymph. Annals of Surgery.1957; 146: 596–06.
    20.  Lee ER. Electromagnetic superficial heating technology. Thermo radiotherapy and thermos chemotherapy, Springer-Verlag. 1995:193-17.
    21.  James C L. Advances in Electromagnetic Fields in Living Systems. Springer Science Business Media.2005; l4: 59.
    22.  Wust S.Electromagnetic deep heating technology, Thermo radiotherapy and thermo chemotherapy. Springer-Verlag.1995:219-51.
    23.  Linh PH, Thach PV, Tuan NA, Thuan NC Manh DH, Phuc NX. Magnetic fluid based on Fe3O4 nanoparticles: Preparation and hyperthermia application. Journal of Physics. 2009; 187: 1-5.
    24.  Delaney G, Jacob S, Featherstone C, Barton M. The role of radiotherapy in cancer treatment: estimating optimal utilization from a review of evidence-based clinical guidelines. Cancer.2005; 104: 1129–37.
    25.  Schaue D, Bride WH. Opportunities and challenges of radiotherapy for treating cancer.Nat. Rev.Clin. Oncol. 2015;12:527–40.
    26.  Vaishnava PP. Magnetic relaxation and dissipative heating in ferrofluids. Journal of Applied Physics. 2007; 102.
    27.  Katz E,Willner I. Integrated nanoparticle-biomolecule hybrid systems: synthesis, properties, and applications. Angewandte Chemie.2004;43(45): 6042–08.
    28.  Ali LO. Study effect of breast cancer on some hematological and biological parameters in Babylon province. Iraq Journal Pharmacy and Biological Sciences. 2014; 9: 20-24.
    29.  Ufelle SA, Ukaejiofo EO, Neboh EE, Achukwu P U, Ikekpeazu E J, Maduka I C, et al. Some haematological parameters in pre-and post- surgery breast cancer patients in Emugu, Nigeria. Int J Cur Bio Med Sci. 2012; 2: 188-90.
    30. Abdelhalim MA, Moussa SA. The dimensional hematological alteration induced in blood of rats in vivo by intraperitoneal administration of gold nanoparticles. Nanomed and Nanotechnol.2012; 3:138.
    31. JoMR, BaeSH, GoMR, KimHJ, HwangYG, hoiSJ. Toxicity and biokinetics of colloidal gold nanoparticles. Nanomaterials.2015; 5:835-50.
    32. Palacin PC, Hidber J,Bourgoin C,Miramond C, Fermon and G. Whitesides. Patterning with magnetic materials at the micron scale. Chem.Mater.1996; 8:1316-25.
    33.  Enzel p, Adelman NB, Beckman KJ, Campbell DJ, Ellis AB, Lisensky GC. Preparation and Properties of an Aqueous Ferrofluid. J. Chem. Educ. 1999;76: 943.
    34.  Linh PH, Van Thach P, Tuan NA, Thuan NC, Phuc NX. Magnetic fluid based on Fe3O4 nanoparticles: Preparation and hyperthermia application. InJournal of Physics: Conference Series 2009 Sep 1; 187(1): 012069.
    35. Jaganathan SK, Mondhe D, Wani ZA, Pal HC, Mandal M. Effect of honey and eugenol on Ehrlich ascites and solid carcinoma. Journal of Biomedicine and Biotechnology. 2010 Oct;2010.
    36. Reilly MS, Boehm T, Shing Y. Endostatin: an endogenous inhibitor of angiogenesis and tumor growth. Cell.1997;88(2):277‐85.
    37.  Mohamed NA, Ahmed MK. Comparative study between the effect of methotrexate and valproic acid on solid Ehrlich tumour. Journal of the Egyptian National Cancer Institute.2012; 24(4):161-7.
    38.  Mohamed MM, UchidaT, Minami S. A plulse operated microwave induced plasma source. Spectrosc. 1989;43: 129 37.
    39.  Begg C, Stewart A, Vens C. Strategies to improve radiotherapy with targeted drugs. Nat Rev Cancer. 2011; 11:239–53.
    40.  Rajamanickam B, Kuo L, Richard Y and KhengW. Cancer and Radiation Therapy: Current Advances and Future Directions. Int. J. Med. Sci. 2012; 9(3):193-9.
    41.  Hanna S. RF Linear Accelerators for Medical and Industrial Applications. Artech House Publishing. 2012.
    42.  Podgorsak EB. Radiation Oncology Physics: A Handbook for Teachers and Students.Vienna: International Atomic Energy Agency (IAEA);2005.
    43.  Ramnik S. Medical Laboratory Technology: methods and Interpretations. 5th ed. Jayee Brothers. New Delhi. 1999.
    44.  Gao ZX, Wang WM, Abbas K, Zhou XY, Yang Y,Diana JS, Wang H P, Wang HL, Li Y, Sun YH. Haematological characterization of loach Misgurnus anguillicaudatus: comparison among diploid, triploid and tetraploid specimens. Comparative Biochemistry and Physiology, Part A: Molecular &Integrative Physiology. 2007a;147(4):1001–08.
    45.  Gao ZX, Wang WM, Yang Y, Abbas K, Li DP, Zuo GW, Diana JS. Morphological studies of peripheral blood cells of the Chinese sturgeon,Acipenser sinensis. Fish Physiology and Biochemistry.2007b; 33:213–22.
    46.  Cao XY, Wang WM. Haematological and biochemical characteristics of two aquacultured carnivorous cyprinids, topmouth culter Culter alburnus (Basilewsky) and yellowcheek carp Elopichthys bambusa (Richardson). Aquaculture Research. 2010; 41: 1331–38.
    47.  Reitman S, Frankel S. Glutamic – Pyruvate Transaminase Assay by Colorimetric Method. Am. J. Clin. 1957; Path 28: 56.       
    48.  Ohkawa H, Ohishi N, Yagi K. Assay of lipid peroxides in animal tissues by thiobarbituric acid reaction. Annals of Biochemistry. 1979; (95):351-58.
    49.  Zettner A. Seligson. Application of atomic absorption spectrophotometry in the determination of calcium in serum. Clin. Chem. 1964;10: 869-89.
    50.  Su B, Xiang SL, Su J. Diallyl disulfide increases histone acetylation and P21WAF1 expression in human gastric cancer cells in vivo and in vitro. Biochemical Pharmacology. 2012; 1(7):1–10.
    51. Siegel L, Miller D, Jemal A. Cancer statistics. CA Cancer J Clin. 2016; 66:7–30.    
    52. Jiajiang X, Zhongxiong F, Yang l, Yinying Z, Fei Y, guanghao S, et al. Design of ph-sensitive methotrexate prodrug-targeted curcumin nanoparticles for efficient dual-drug delivery and combination cancer therapy. International Journal of Nanomedicine. 2018; 13:1381–98.
    53. Brigger I, Dubernet C, Couvreur P. Nanoparticles in cancer therapy and diagnosis. Adv Drug Deliv Rev. 2002; 54:631–51.
    54. Wu Q, Yang Z, Nie Y, Shi Y, Fan D. Multi-drug resistance in cancer chemotherapeutics: mechanisms and lab approaches. Cancer Lett. 2014; 347:159–66.
    55. Lehár J, Krueger AS, Avery W, Heilbut AM, Johansen LM, Price ER, et al. Synergistic drug combinations tend to improve therapeutically relevant selectivity. Nature biotechnology. 2009 Jul;27(7):659-66.
    56. Liu Z, Jiao Y, Wang Y, Zhou C, Zhang Z. Polysaccharides-based nanoparticles as drug delivery systems. Adv Drug Deliv Rev. 2008; 60:1650–62.
    57. Song L, Pan Z, Zhang H, et al. Dually folate/Cd44 receptor-targeted self-assembled hyaluronic acid nanoparticles for dual-drug delivery and combination cancer therapy. J Mater Chem B. 2017; 5:6835–46.
    58.  Thomas TP, Huang B, Choi SK, Silpe JE, Kotlyar A, Desai AM, et al. Polyvalent dendrimer-methotrexate as a folate receptor-targeted cancer therapeutic. Molecular pharmaceutics. 2012 Sep 4;9(9):2669-76.
    59. Zeb A, Qureshi OS, Kim HS, Cha JH, Kim HS, Kim JK. Improved skin permeation of methotrexate via nanosized ultradeformable liposomes. Int J Nanomedicine. 2016;11:3813–24.
    60.  Boechat AL, de Oliveira CP, Tarragô AM, da Costa AG, Malheiro A, Guterres SS, et al. Methotrexate-loaded lipid-core nanocapsules are highly effective in the control of inflammation in synovial cells and a chronic arthritis model. International journal of nanomedicine. 2015;10:6603.
    61. Mount C, Featherstone J. Rheumatoid arthritis market. Nat Rev Drug Discov. 2005;4:11–12.
    62. Shao Z, Shao J, Tan B, Guan S, Liu Z, Zhao Z, et al. Targeted lung cancer therapy: preparation and optimization of transferrin-decorated nanostructured lipid carriers as novel nanomedicine for co-delivery of anticancer drugs and DNA. International journal of nanomedicine. 2015; 10:1223.
    63. Rex DK, Kahi CJ, Levin B, Smith RA, Bond JH, Brooks D. Guidelines for colonoscopy surveillance after cancer resection: a consensus update by the American Cancer Society and the US Multi-Society Task Force on Colorectal Cancer. Gastroenterology. 2006; 130(6):1865-71. DOI: 10.1053/j.gastro.2006.03.013
    64. Bonvin D, et al. Folic acid on iron oxide nanoparticles: platform with high potential for simultaneous targeting, MRI detection and hyperthermia treatment of lymph node metastases of prostate cancer. Dalton Trans. 2017;46(37):12692–704.
    65.  Peer D, Karp JM, Hong S, Farokhzad OC, Margalit R, Langer R. Nanocarriers as an emerging platform for cancer therapy. Nature nanotechnology. 2007 Dec;2(12):751-60.
    66.  Wang X, Yang L, Chen Z, Shin DM. Application of nanotechnology in cancer therapy and imaging, CA: Cancer J. Clin. 2008; 58: 97–110.
    67.  Fang J, Nakamura H, Maeda H, The EPR effect: Unique features of tumor blood vessels for drug delivery, factors involved, and limitations and augmentation of the effect. Adv. Drug Deliv. 2011; 63:136–51.
    68.  Gong J, Chen M, Zheng Y, Wang S, Wang Y. Polymeric micelles drug delivery system in oncology. J. Control. Release. 2012; 159 :312–23.
    69.  Cheng R, Meng F, Deng C, Klok H-A, Zhong Z. Dual and multi-stimuli responsive polymeric nanoparticles for programmed site-specific drug delivery. Biomaterials. 2013; 34: 3647–57.
    70.  Nedkov T, Merodiiska L, Slavov RE, Vandenberghe Y,Kusano J, Takada J. Surface oxidation, size and shape of nano-sized magnetite obtained by co-precipitation. Magn. And Magn. Mater. 2006 ;300(2): 358-67.
    71.  Cengelli F. Interaction of functionalization superparamagnetic iron oxide nanoparticles with brain structures. Journal Of Pharmacology And Experimental Therapeutics. 2006; 318: 108-16. DOI:10.1124/ipet.106.101915.
    72. Rasha SS, Mohamed MM, Tayel MB, Ali NH. Effect of microwave hyperthermia on tumor treatment. Romanian J. Biophys. 2013; 23(4): 249-59