Advertisement

Improving the Subcutaneous Mouse Tumor Model by Effective Manipulation of Magnetic Nanoparticles-Treated Implanted Cancer Cells

  • Katerina Spyridopoulou
  • Georgios Aindelis
  • Evangeli Lampri
  • Maria Giorgalli
  • Eleftheria Lamprianidou
  • Ioannis Kotsianidis
  • Anastasia Tsingotjidou
  • Aglaia Pappa
  • Orestis Kalogirou
  • Katerina Chlichlia
Article

Abstract

Murine tumor models have played a fundamental role in the development of novel therapeutic interventions and are currently widely used in translational research. Specifically, strategies that aim at reducing inter-animal variability of tumor size in transplantable mouse tumor models are of particular importance. In our approach, we used magnetic nanoparticles to label and manipulate colon cancer cells for the improvement of the standard syngeneic subcutaneous mouse tumor model. Following subcutaneous injection on the scruff of the neck, magnetically-tagged implanted cancer cells were manipulated by applying an external magnetic field towards localized tumor formation. Our data provide evidence that this approach can facilitate the formation of localized tumors of similar shape, reducing thereby the tumor size’s variability. For validating the proof-of-principle, a low-dose of 5-FU was administered in small animal groups as a representative anticancer therapy. Under these experimental conditions, the 5-FU-induced tumor growth inhibition was statistically significant only after the implementation of the proposed method. The presented approach is a promising strategy for studying accurately therapeutic interventions in subcutaneous experimental solid tumor models allowing for the detection of statistically significant differences between smaller experimental groups.

Keywords

Magnetic nanoparticles Subcutaneous/transplantable mouse tumor model BALB/c mice CT26 cells Preclinical screening 

Abbreviations

5-FU

Fluorouracil

DMEM

Dulbecco’s modified Eagle medium

DNA

Deoxyribonucleic acid

ICP-OES

Inductively coupled plasma optical emission spectrometry

PBS

Phosphate-buffered saline

SRB

Sulforhodamine B

SSC

Side scatter

XTT

2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide

MNPs

Magnetic nanoparticles

flMNPs

Fluorescently-labeled magnetic nanoparticles

CI

Confidence interval

SD

Standard deviation

IQR

Interquartile range

Notes

Acknowledgments

Part of the work was implemented by utilizing the facilities of the ‘OPENSCREEN-GR’ supported by the National Roadmap for Research Infrastructures under the National Strategy for Research, Technological Development, and Innovation (2014–2020) by the General Secretariat for Research and Technology (GSRT), Ministry of Education and Religious Affairs, Hellenic Republic.

Conflict of interest

Authors have no conflicts of interest to declare.

References

  1. 1.
    Alon, N., T. Havdala, H. Skaat, K. Baranes, M. Marcus, I. Levy, S. Margel, A. Sharoni, and O. Shefi. Magnetic micro-device for manipulating PC12 cell migration and organization. Lab Chip 15:2030–2036, 2015.CrossRefPubMedGoogle Scholar
  2. 2.
    Bradshaw, M., T. D. Clemons, D. Ho, L. Gutiérrez, F. J. Lázaro, M. J. House, T. G. St. Pierre, M. W. Fear, F. M. Wood, and K. S. Iyer. Manipulating directional cell motility using intracellular superparamagnetic nanoparticles. Nanoscale 7:4884–4889, 2015.CrossRefPubMedGoogle Scholar
  3. 3.
    Castle, J. C., M. Loewer, S. Boegel, J. de Graaf, C. Bender, A. D. Tadmor, V. Boisguerin, T. Bukur, P. Sorn, C. Paret, M. Diken, S. Kreiter, Ö. Türeci, and U. Sahin. Immunomic, genomic and transcriptomic characterization of CT26 colorectal carcinoma. BMC Genomics 15:190, 2014.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Chen, J., N. Huang, B. Ma, M. F. Maitz, J. Wang, J. Li, Q. Li, Y. Zhao, K. Xiong, and X. Liu. Guidance of stem cells to a target destination in vivo by magnetic nanoparticles in a magnetic field. ACS Appl. Mater. Interfaces 5:5976–5985, 2013.CrossRefPubMedGoogle Scholar
  5. 5.
    Day, C. P., G. Merlino, and T. Van Dyke. Preclinical mouse cancer models: a maze of opportunities and challenges. Cell 163:39–53, 2015.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    De Jong, M., and T. Maina. Of mice and humans: are they the same? Implications in cancer translational research. J. Nucl. Med. 51:501–504, 2010.CrossRefPubMedGoogle Scholar
  7. 7.
    Dranoff, G. Experimental mouse tumour models: what can be learnt about human cancer immunology? Nat. Rev. Immunol. 12:61–66, 2012.CrossRefGoogle Scholar
  8. 8.
    Farkona, S., E. P. Diamandis, and I. M. Blasutig. Cancer immunotherapy: the beginning of the end of cancer? BMC Med. 2016.  https://doi.org/10.1186/s12916-016-0623-5.PubMedPubMedCentralGoogle Scholar
  9. 9.
    Friedrich, R. P., C. Janko, M. Poettler, P. Tripal, J. Zaloga, I. Cicha, S. Dürr, J. Nowak, S. Odenbach, I. Slabu, M. Liebl, L. Trahms, M. Stapf, I. Hilger, S. Lyer, and C. Alexiou. Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods. Int. J. Nanomed. 10:4185, 2015.CrossRefGoogle Scholar
  10. 10.
    Haddad, T. C., and D. Yee. Of mice and (wo)men: is this any way to test a new drug? J. Clin. Oncol. 26:830–832, 2008.CrossRefPubMedGoogle Scholar
  11. 11.
    Hughes, C. S., L. M. Postovit, and G. A. Lajoie. Matrigel: a complex protein mixture required for optimal growth of cell culture. Proteomics 10:1886–1890, 2010.CrossRefPubMedGoogle Scholar
  12. 12.
    Kasten, A., C. Grüttner, J.-P. Kühn, R. Bader, J. Pasold, and B. Frerich. Comparative in vitro study on magnetic iron oxide nanoparticles for MRI tracking of adipose tissue-derived progenitor cells. PLoS ONE 9:e108055, 2014.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Kim, J. A., C. Åberg, A. Salvati, and K. A. Dawson. Role of cell cycle on the cellular uptake and dilution of nanoparticles in a cell population. Nat. Nanotechnol. 7:62–68, 2011.CrossRefPubMedGoogle Scholar
  14. 14.
    Kolosnjaj-Tabi, J., C. Wilhelm, O. Clément, and F. Gazeau. Cell labeling with magnetic nanoparticles: opportunity for magnetic cell imaging and cell manipulation. J. Nanobiotechnology 11:S7, 2013.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Lechner, M. G., S. S. Karimi, K. Barry-holson, and T. E. Angell. NIH public access. J. Immunother. 36:477–489, 2014.CrossRefGoogle Scholar
  16. 16.
    Liu, J., X. Tian, M. Bao, J. Li, Y. Dou, B. Yuan, K. Yang, and Y. Ma. Manipulation of cellular orientation and migration by internalized magnetic particles. Mater. Chem. Front. Mater. Chem. Front 1:933–936, 2017.CrossRefGoogle Scholar
  17. 17.
    Longley, D. B., D. P. Harkin, and P. G. Johnston. 5-Fluorouracil: mechanisms of action and clinical strategies. Nat. Rev. Cancer 3:330–338, 2003.CrossRefPubMedGoogle Scholar
  18. 18.
    Milotti, E., V. Vyshemirsky, M. Sega, and R. Chignola. Interplay between distribution of live cells and growth dynamics of solid tumours. Sci. Rep. 2:1–10, 2012.CrossRefGoogle Scholar
  19. 19.
    Qian, L., Y. Liu, S. Wang, W. Gong, X. Jia, L. Liu, F. Ye, J. Ding, Y. Xu, Y. Fu, and F. Tian. NKG2D ligand RAE1ε induces generation and enhances the inhibitor function of myeloid-derived suppressor cells in mice. J. Cell Mol. Med. 21:2046–2054, 2017.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Rossi, L., E. Laas, P. Mallon, A. Vincent-Salomon, J. M. Guinebretiere, F. Lerebours, R. Rouzier, J. Y. Pierga, and F. Reyal. Prognostic impact of discrepant Ki67 and mitotic index on hormone receptor-positive, HER2-negative breast carcinoma. Br. J. Cancer 113:996–1002, 2015.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Sagiv-Barfi, I., D. K. Czerwinski, S. Levy, I. S. Alam, A. T. Mayer, S. S. Gambhir, and R. Levy. Eradication of spontaneous malignancy by local immunotherapy. Sci. Transl. Med. 2018.  https://doi.org/10.1126/scitranslmed.aan4488.PubMedPubMedCentralGoogle Scholar
  22. 22.
    Schleich, N., P. Sibret, P. Danhier, B. Ucakar, S. Laurent, R. N. Muller, C. Jérôme, B. Gallez, V. Préat, and F. Danhier. Dual anticancer drug/superparamagnetic iron oxide-loaded PLGA-based nanoparticles for cancer therapy and magnetic resonance imaging. Int. J. Pharm. 447:94–101, 2013.CrossRefPubMedGoogle Scholar
  23. 23.
    Seeliger, H., M. Guba, G. E. Koehl, A. Doenecke, M. Steinbauer, C. J. Bruns, C. Wagner, E. Frank, K. W. Jauch, and E. K. Geissler. Blockage of 2-deoxy-D-ribose-induced angiogenesis with rapamycin counteracts a thymidine phosphorylase-based escape mechanism available for colon cancer under 5-fluorouracil therapy. Clin. Cancer Res. 10:1843–1852, 2004.CrossRefPubMedGoogle Scholar
  24. 24.
    Silva, L. H. A., F. F. Cruz, M. M. Morales, D. J. Weiss, and P. R. M. Rocco. Magnetic targeting as a strategy to enhance therapeutic effects of mesenchymal stromal cells. Stem Cell Res. Ther. 8:58, 2017.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Spyridopoulou, K., A. Makridis, N. Maniotis, N. Karypidou, E. Myrovali, T. Samaras, M. Angelakeris, K. Chlichlia, and O. Kalogirou. Effect of low frequency magnetic fields on the growth of MNPs-treated HT29 colon cancer cells. Nanotechnology 29:175101, 2018.CrossRefPubMedGoogle Scholar
  26. 26.
    Spyridopoulou, K., A. Tiptiri-Kourpeti, E. Lampri, E. Fitsiou, S. Vasileiadis, M. Vamvakias, H. Bardouki, A. Goussia, V. Malamou-Mitsi, M. I. Panayiotidis, A. Galanis, A. Pappa, and K. Chlichlia. Dietary mastic oil extracted from Pistacia lentiscus var. chia suppresses tumor growth in experimental colon cancer models. Sci. Rep. 7:3782, 2017.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Suggitt, M., and M. C. Bibby. 50 years of preclinical anticancer drug screening : empirical to target-driven approaches. Clin. Cancer Res. 11:971–981, 2005.PubMedGoogle Scholar
  28. 28.
    Sullivan, K. M., A. Dean, and M. S. Minn. OpenEpi: a web-based epidemiologic and statistical calculator for public health. Public Health Rep. 124:471–474, 2009.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Summers, H. Bionanoscience: nanoparticles in the life of a cell. Nat. Nanotechnol. 7:9–10, 2011.CrossRefPubMedGoogle Scholar
  30. 30.
    Suzuki, H., T. Toyooka, and Y. Ibuki. Simple and easy method to evaluate uptake potential of nanoparticles in mammalian cells using a flow cytometric light scatter analysis. Environ. Sci. Technol. 41:3018–3024, 2007.CrossRefPubMedGoogle Scholar
  31. 31.
    Talmadge, J. E., R. K. Singh, I. J. Fidler, and A. Raz. Murine models to evaluate novel and conventional therapeutic strategies for cancer. Am. J. Pathol. 170:793–804, 2007.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Theumer, A., C. Gräfe, F. Bähring, C. Bergemann, A. Hochhaus, and J. H. Clement. Superparamagnetic iron oxide nanoparticles exert different cytotoxic effects on cells grown in monolayer cell culture versus as multicellular spheroids. J. Magn. Magn. Mater. 380:27–33, 2015.CrossRefGoogle Scholar
  33. 33.
    Wagner, M., V. Roh, M. Strehlen, A. Laemmle, D. Stroka, B. Egger, M. Trochsler, K. K. Hunt, D. Candinas, and S. A. Vorburger. Effective treatment of advanced colorectal cancer by rapamycin and 5-FU/oxaliplatin monitored by TIMP-1. J. Gastrointest. Surg. 13:1781–1790, 2009.CrossRefPubMedGoogle Scholar
  34. 34.
    Weissleder, R., M. Nahrendorf, and M. J. Pittet. Imaging macrophages with nanoparticles. Nat. Mater. 13:125–138, 2014.CrossRefPubMedGoogle Scholar
  35. 35.
    Whiteside, T. The tumor microenvironment and its role in promoting tumor growth. Oncogene 27:5904–5912, 2013.CrossRefGoogle Scholar
  36. 36.
    Wilhelm, C., F. Gazeau, J. Roger, J. N. Pons, and J. C. Bacri. Interaction of anionic superparamagnetic nanoparticles with cells: kinetic analyses of membrane adsorption and subsequent internalization. Langmuir 18:8148–8155, 2002.CrossRefGoogle Scholar
  37. 37.
    Yanai, A., U. O. Häfeli, A. L. Metcalfe, P. Soema, L. Addo, C. Y. Gregory-Evans, K. Po, X. Shan, O. L. Moritz, and K. Gregory-Evans. Focused magnetic stem cell targeting to the retina using superparamagnetic iron oxide nanoparticles. Cell Transplant. 21:1137–1148, 2012.CrossRefPubMedGoogle Scholar
  38. 38.
    Zitvogel, L., J. M. Pitt, R. Daillère, M. J. Smyth, and G. Kroemer. Mouse models in oncoimmunology. Nat. Rev. Cancer 16:759–773, 2016.CrossRefPubMedGoogle Scholar

Copyright information

© Biomedical Engineering Society 2018

Authors and Affiliations

  • Katerina Spyridopoulou
    • 1
  • Georgios Aindelis
    • 1
  • Evangeli Lampri
    • 1
  • Maria Giorgalli
    • 1
  • Eleftheria Lamprianidou
    • 2
  • Ioannis Kotsianidis
    • 2
  • Anastasia Tsingotjidou
    • 3
  • Aglaia Pappa
    • 1
  • Orestis Kalogirou
    • 4
  • Katerina Chlichlia
    • 1
  1. 1.Department of Molecular Biology and GeneticsDemocritus University of ThraceAlexandroupolisGreece
  2. 2.Department of Hematology, School of MedicineDemocritus University of ThraceAlexandroupolisGreece
  3. 3.Laboratory of Anatomy, Histology and Embryology, School of Veterinary MedicineAristotle University of ThessalonikiThessalonikiGreece
  4. 4.Department of PhysicsAristotle University of ThessalonikiThessalonikiGreece

Personalised recommendations