Advertisement

Biomedical Microdevices

, 21:94 | Cite as

Investigation of uniform sized multicellular spheroids raised by microwell arrays after the combined treatment of electric field and anti-cancer drug

  • Kin Fong LeiEmail author
  • Wun-Wu Ji
  • Andrew Goh
  • Chun-Hao Huang
  • Ming-Yih LeeEmail author
Article
  • 29 Downloads

Abstract

Nowadays, cancer disease is continuously identified as the leading cause of mortality worldwide. Cancer chemotherapeutic agents have been continuously developing to achieve high curative effectiveness and low side effects. However, solid tumors present the properties of low drug penetration and resistance of quiescent cells. Radiation therapy is concurrently given in some cases; but it induces different levels of adverse effects. In the current work, uniform sized multicellular spheroids were raised by microwell arrays to mimic the architecture of solid tumors. Investigation of the response of the spheroids was conducted after the treatment of alternating electric field. The result showed that the electric field could induce early apoptosis by disturbing cell membrane. Moreover, combined treatment of electric field and anti-cancer drug was applied to the spheroids. The electric field synergistically enhanced the treatment efficacy because the anti-cancer drug could permeate through the disrupted cell membrane. Significant improvement of late apoptosis was shown by the combined treatment. Because the electric field treatment induces limited side effect to the patient, lower dosage of anti-cancer drug may be applied to the patients for achieving curative effectiveness.

Keywords

Multicellular spheroids Microwell arrays Electric field therapy Cell apoptosis 

Notes

Acknowledgements

This work was supported by Chang Gung Memorial Hospital, Linkou, Taiwan (project no. CMRPD2H0022 and BMRPC05).

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest.

Supplementary material

10544_2019_442_MOESM1_ESM.pdf (928 kb)
ESM 1 (PDF 928 kb)

References

  1. N.H. Baek, O.W. Seo, M.S. Kim, J. Hulme, S.S.A. An, Monitoring the effects of doxorubicin on 3D-spheroid tumor cells in real-time. Oncotargets Ther. 9, 7207–7218 (2016)CrossRefGoogle Scholar
  2. G.C. Barnett, C.M.L. West, A.M. Dunning, R.M. Elliott, C.E. Coles, et al., Normal tissue reactions to radiotherapy: Towards tailoring treatment dose by genotype. Nature Rev. 9, 134–142 (2009)Google Scholar
  3. K.K. Brown, L. Montaser-Kouhsari, A.H. Beck, A. Toker, MERIT40 is a AKT substrate that promotes resolution of DNA damage induced by chemotherapy. Cell Rep. 11, 1358–1366 (2015)CrossRefGoogle Scholar
  4. D.N. Carney, J.B. Mitchell, T.J. Kinsella, In vitro radiation and chemotherapy sensitivity of established cell lines of human small cell lung cancer and its large cell morphological variants. Cancer Res. 43, 2806–2811 (1983)Google Scholar
  5. M. Charnley, M. Textor, A. Khademhosseini, M.P. Lutolf, Integration column: Microwell arrays for mammalian cell culture. Integr. Biol. 1(11–12), 625–634 (2009)CrossRefGoogle Scholar
  6. Y.C. Chen, X. Lou, Z. Zhang, P. Ingram, E. Yoon, High-throughput cancer cell sphere formation for characterizing the efficacy of photo dynamic therapy in 3D cell cultures. Sci. Rep. 5, 12175 (2015)CrossRefGoogle Scholar
  7. E.C. Costa, V.M. Gaspar, P. Coutinho, I.J. Correia, Optimization of liquid overlay technique to formulate heterogenic 3D co-culture models. Biotechnol. Bioeng. 111, 1672–1685 (2014)CrossRefGoogle Scholar
  8. C. Dubessy, J.M. Merlin, C. Marchal, F. Guillemin, Spheroids in radiobiology and photodynamic therapy. Crit Rev Oncol Hematol 36(2–3), 179–192 (2000)CrossRefGoogle Scholar
  9. E. Fennema, N. Rivron, J. Rouwkema, C. van Blitterswijk, J. de Boer, Spheroid culture as a tool for creating 3D complex tissues. Trends Biotechnol. 31, 108–115 (2013)CrossRefGoogle Scholar
  10. J. Friedrich, R. Ebner, L.A. Kunz-Schughart, Experimental anti-tumor therapy in 3-D: Spheroids–old hat or new challenge? Int. J. Radiat. Biol. 83(11–12), 849–871 (2007)CrossRefGoogle Scholar
  11. N. Gera, N., A. Yang, T.S. Holtzman, S.X. Lee, E.T. Wong, et al., Tumor treating fields perturb the localization of septins and cause aberrant mitotic exit. PLoS One 2015, 10, e125269CrossRefGoogle Scholar
  12. M. Giladi, U. Weinberg, R.S. Schneiderman, Y. Porat, M. Munster, et al., Alternating electric fields (tumor-treating fields therapy) can improve chemotherapy treatment efficacy in non-small cell lung cancer both in vitro and in vivo. Semin. Oncol. 41, S35–S41 (2014)CrossRefGoogle Scholar
  13. M. Giladi, R.S. Schneiderman, T. Voloshin, Y. Porat, M. Munster, et al., Mitotic spindle disruption by alternating electric fields leads to improper chromosome segregation and mitotic catastrophe in cancer cells. Sci. Rep. 5, 18046 (2015)CrossRefGoogle Scholar
  14. X. Gong, C. Lin, J. Cheng, J. Su, H. Zhao, T. Liu, X. Wen, P. Zhao, Generation of multicellular tumor spheroids with microwell-based agarose scaffolds for drug testing. PLoS One 10(6), e0130348 (2015)CrossRefGoogle Scholar
  15. F. Hirschhaeuser, H. Menne, C. Dittfeld, J. West, W. Mueller-Klieser, L.A. Kunz-Schughart, Multicellular tumor spheroids: An underestimated catching up again. J. Biotechnol. 148(1), 3–15 (2010)CrossRefGoogle Scholar
  16. C. Isanbor, D. O’Hagan, Fluorine in medicinal chemistry: A review of anti-cancer agents. J. Fluor. Chem. 127, 303–319 (2006)CrossRefGoogle Scholar
  17. E.H. Kim, Y.J. Kim, H.S. Song, Y.K. Jeong, J.Y. Lee, et al., Biological effect of an alternating electric field on cell proliferation and synergistic antimitotic effect in combination with ionizing radiation. Oncotarget 7, 62267–62279 (2016a)Google Scholar
  18. E.H. Kim, H.S. Song, S.H. Yoo, M. Yoon, Tumor treating fields inhibit glioblastoma cell migration, invasion and angiogenesis. Oncotarget 7, 65125–65136 (2016b)Google Scholar
  19. E.D. Kirson, Z. Gurvich, R. Schneiderman, E. Dekel, A. Itzhaki, et al., Disruption of Cancer cell replication by alternating electric fields. Cancer Res. 64, 3288–3295 (2004)CrossRefGoogle Scholar
  20. E.D. Kirson, V. Dbaly, F. Tovarys, J. Vymazal, J.F. Soustiel, et al., Alternating electric fields arrest cell proliferation in animal tumor models and human brain tumors. Proc. Natl. Acad. Sci. U. S. A. 104, 10152–10157 (2007)CrossRefGoogle Scholar
  21. E.D. Kirson, M. Giladi, Z. Gurvich, A. Itzhaki, D. Mordechovich, et al., Alternating electric fields (TTFields) inhibit metastatic spread of solid tumors to the lungs. Clin Exp Metastas 26, 633–640 (2009)CrossRefGoogle Scholar
  22. V. Koshkin, L.E. Ailes, G. Liu, S.N. Krylov, Metabolic suppression of a drug-resistant subpopulation in cancer spheroid cells. J. Cell. Biochem. 117, 59–65 (2016)CrossRefGoogle Scholar
  23. J. Laurent, C. Frongia, M. Cazales, O. Mondesert, B. Ducommun, V. Lobjois, Multicellular tumor spheroid models to explore cell cycle checkpoints in 3D. BMC Cancer 13, 73 (2013)CrossRefGoogle Scholar
  24. R.Z. Lin, H.Y. Chang, Recent advances in three-dimensional multicellular spheroid culture for biomedical research. Biotechnol. J. 3, 1172–1184 (2008)CrossRefGoogle Scholar
  25. F.F. Liu, C. Peng, B.I. Escher, E. Fantino, C. Giles, S. Were, L. Duffy, J.C. Ng, Hanging drop: An in vitro air toxic exposure model using human lung cells in 2D and 3D structures. J. Hazard. Mater. 261, 701–710 (2013)CrossRefGoogle Scholar
  26. P. Longati, X. Jia, J. Eimer, A. Wagman, M.R. Witt, et al., 3D pancreatic carcinoma spheroids induce a matrix-rich, chemoresistant phenotype offering a better model for drug testing. BMC Cancer 13, 95 (2013) (13pp)CrossRefGoogle Scholar
  27. F. Pampaloni, E.G. Reynaud, E.H.K. Stelzer, The third dimension bridges the gap between cell culture and live tissue. Nat Rev Mole Cell Bio 8, 839–845 (2007)CrossRefGoogle Scholar
  28. F. Perche, V.P. Torchilin, Cancer cell spheroids as a model to evaluate chemotherapy protocols. Cancer Biol Ther 13, 1205–1213 (2012)CrossRefGoogle Scholar
  29. M. Pless, C. Drogege, R. von Moss, M. Salzberg, D. Betticher, A phase I/II trial of tumor treating fields (TTFields) therapy in combination with pemetrexed for advanced non-small cell lung cancer. Lung Cancer 81, 445–450 (2013)CrossRefGoogle Scholar
  30. S. Raghavan, M.R. Ward, K.R. Rowley, R.M. Wold, S. Takayama, R.J. Buckanovich, G. Mehta, Formation of stable small cell number three-dimensional ovarian cancer spheroids using hanging drop arrays for preclinical drug sensitivity assays. Gynecol. Oncol. 138, 181–189 (2015)CrossRefGoogle Scholar
  31. A. Seyfoori, E. Samiei, N. Jalili, B. Godau, M. Rahmanian, L. Farahmand, K. Majidzadeh-A, M. Akbari, Self-filling microwell arrays (SFMAs) for tumor spheroid formation. Lab Chip 18, 3516–3528 (2018)CrossRefGoogle Scholar
  32. H.B. Stone, C.N. Coleman, M.S. Anscher, W.H. McBride, Effects of radiation on normal tissue: Consequences and mechanisms. Lancet Oncol. 4, 529–536 (2003)CrossRefGoogle Scholar
  33. R. Stupp, S. Taillbert, A.A. Kanner, S. Kesari, D.M. Steinberg, et al., Maintenance therapy with tumor-treating fields plus temozolomide vs temozolomide alone for glioblastoma. JAMA 314, 2535–2543 (2015)CrossRefGoogle Scholar
  34. R. Stupp, S. Taillibert, A.A. Kanner, W. Read, D.M. Steinberg, et al., Effect of tumor-treating fields plus maintenance temozolomide vs maintenance temozolomide alone on survival in patients with glioblastoma. JAMA 318, 2306–2316 (2017)CrossRefGoogle Scholar
  35. R.M. Sutherland, Cell and environment interactions in tumor microregions: The multicell spheroid model. Science 240(4849), 177–184 (1988)CrossRefGoogle Scholar
  36. O. Tacar, P. Sriamornask, C.R. Dass, Doxorubicin: An update on anticancer molecular action, toxicity and novel drug delivery systems. J. Pharm. Pharmacol. 65, 157–170 (2013)CrossRefGoogle Scholar
  37. Y.C. Tung, A.Y. Hsiao, S.G. Allen, Y.S. Torisawa, M. Ho, S. Takayama, High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array. Analyst 136, 473–478 (2011)CrossRefGoogle Scholar
  38. J.L. Villano, L.E. Williams, K.S. Watson, N. Ignatius, M.T. Wilson, et al., Delayed response and survival from NovoTTF-100A in recurrent GBM. Med. Oncol. 30, 338 (2013)CrossRefGoogle Scholar
  39. J. Vymazal, E.T. Wong, Response patterns of recurrent glioblastomas treated with tumor-treating fields. Semin. Oncol. 41, S14–S24 (2014)CrossRefGoogle Scholar
  40. E.T. Wong, E. Lok, K.D. Swanson, S. Gautam, H.H. Engelhard, et al., Response assessment of NovoTTF-100A versus best physician’s choice chemotherapy in recurrent glioblastoma. Cancer Med 3, 592–602 (2014)CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Graduate Institute of Biomedical EngineeringChang Gung UniversityTao-YuanTaiwan
  2. 2.Department of Radiation OncologyChang Gung Memorial HospitalLinkouTaiwan
  3. 3.PhD Program in Biomedical Engineering, College of EngineeringChang Gung UniversityTaoyuanTaiwan
  4. 4.Department of CardiologyChang Gung Memorial HospitalLinkouTaiwan

Personalised recommendations