Impact of multicellular tumor spheroids as an in vivo‑like tumor model on anticancer drug response

Galateanu,Bianca; Hudita,Ariana; Negrei,Carolina; Ion,Rodica‑Mariana; Costache,Marieta; Stan,Miriana; Nikitovic,Dragana; Hayes,A. Wallace; Spandidos,Demetrios A.; Tsatsakis,Aristidis M.; Ginghina,Octav

doi:10.3892/ijo.2016.3467

Impact of multicellular tumor spheroids as an in vivo‑like tumor model on anticancer drug response

Authors:
- Bianca Galateanu
- Ariana Hudita
- Carolina Negrei
- Rodica‑Mariana Ion
- Marieta Costache
- Miriana Stan
- Dragana Nikitovic
- A. Wallace Hayes
- Demetrios A. Spandidos
- Aristidis M. Tsatsakis
- Octav Ginghina
View Affiliations

Affiliations: Department of Biochemistry and Molecular Biology, University of Bucharest, Bucharest 050095, Romania, Department of Toxicology, Faculty of Pharmacy, ‘Carol Davila’ University of Medicine and Pharmacy, Bucharest 020956, Romania, National Institute of Research and Development for Chemistry and Petrochemistry ‑ ICECHIM, Bucharest 060021, Romania, Department of Anatomy-Histology-Embryology, Medical School, University of Crete, Heraklion 71003, Greece, Michigan State University, East Lansing, MI 48824, USA, Laboratory of Clinical Virology, University of Crete Medical School, Heraklion 71409, Greece, Department of Forensic Sciences and Toxicology, Medical School, University of Crete, Heraklion 71003, Greece, Department of Surgery, ‘Sf. Ioan’ Emergency Clinical Hospital, ‘Carol Davila’ University of Medicine and Pharmacy, Bucharest 042122, Romania
Published online on: April 1, 2016 https://doi.org/10.3892/ijo.2016.3467
Pages: 2295-2302
Copyright: © Galateanu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The incidence of colorectal cancer is higher in men than in women, amounting to 15% of cancer-related diseases as a whole. As such, undesirable effects, arising from the administration of current chemotherapeutic agents (the FOLFIRI/FOLFOX combinations), which are exerted on the remaining non-cancerous tissues and/or cells, have contributed to the occurrence of resistance to multiple drugs, thus markedly reducing their efficacy. However, the delivery of chemotherapeutic agents may be improved and their action may be more selectively targeted to diseased tissues/cells by means of developing biotechnologies and nano‑techniques. Thus, the current focus is on creating biological tissue and related tumor models, by means of three‑dimensional (3D) spheres, in an attempt to bridge the gap between results obtained in the pre‑clinical phase and promising outcomes obtained in clinical trials. For this purpose, the characterization and use of so‑called ‘multicellular tumor spheroids’, may prove to be invaluable. In this study, we focus on describing the efficacy of a model 3D system as compared to the traditional 2D tumor spheres in determining drug response, highlighting a potentially greater effect of the drugs following the encapsulation of respective liposomes. The results obtained demonstrate the successful preparation of a suspension of liposomes loaded with folinic acid, oxaliplatin and 5‑fluorouracil (5‑FU), and loaded with meso‑tetra (4‑sulfonatophenyl) porphyrin. Following its use on HT‑29 colorectal cancer cells, an important comparative reduction was noted in the viability of the HT‑29 cells, demonstrating the efficacy of multicellular tumor spheroids carrying liposomes loaded with therapeutic drugs. These findings indicate that the method of drug encapsulation in liposomes may improve the treatment efficacy of chemotherapeutic agents.

View Figures

View References

1	Zavoral M, Suchanek S, Zavada F, Dusek L, Muzik J, Seifert B and Fric P: Colorectal cancer screening in Europe. World J Gastroenterol. 15:5907–5915. 2009. View Article : Google Scholar : PubMed/NCBI
2	Jemal A, Clegg LX, Ward E, Ries LA, Wu X, Jamison PM, Wingo PA, Howe HL, Anderson RN and Edwards BK: Annual report to the nation on the status of cancer, 1975–2001 with a special feature regarding survival. Cancer. 101:3–27. 2004. View Article : Google Scholar : PubMed/NCBI
3	Boyle P and Langman JS: ABC of colorectal cancer: Epidemiology. BMJ. 321:805–808. 2000. View Article : Google Scholar : PubMed/NCBI
4	Parkin DM, Bray F, Ferlay J and Pisani P: Global cancer statistics, 2002. CA Cancer J Clin. 55:74–108. 2005. View Article : Google Scholar : PubMed/NCBI
5	Jackson-Thompson J, Ahmed F, German RR, Lai SM and Friedman C: Descriptive epidemiology of colorectal cancer in the United States, 1998–2001. Cancer. 107(Suppl 5): 1103–1111. 2006. View Article : Google Scholar : PubMed/NCBI
6	Mayer RJ: Moving beyond fluorouracil for colorectal cancer. N Engl J Med. 343:963–964. 2000. View Article : Google Scholar : PubMed/NCBI
7	Saltz LB, Cox JV, Blanke C, Rosen LS, Fehrenbacher L, Moore MJ, Maroun JA, Ackland SP, Locker PK, Pirotta N, et al; Irinotecan Study Group. Irinotecan plus fluorouracil and leucovorin for metastatic colorectal cancer. N Engl J Med. 343:905–914. 2000. View Article : Google Scholar : PubMed/NCBI
8	de Gramont A, Figer A, Seymour M, Homerin M, Hmissi A, Cassidy J, Boni C, Cortes-Funes H, Cervantes A, Freyer G, et al: Leucovorin and fluorouracil with or without oxaliplatin as first-line treatment in advanced colorectal cancer. J Clin Oncol. 18:2938–2947. 2000.PubMed/NCBI
9	Giacchetti S, Perpoint B, Zidani R, Le Bail N, Faggiuolo R, Focan C, Chollet P, Llory JF, Letourneau Y, Coudert B, et al: Phase III multicenter randomized trial of oxaliplatin added to chronomodulated fluorouracil-leucovorin as first-line treatment of metastatic colorectal cancer. J Clin Oncol. 18:136–147. 2000.PubMed/NCBI
10	Scheithauer W, Rosen H, Kornek GV, Sebesta C and Depisch D: Randomised comparison of combination chemotherapy plus supportive care with supportive care alone in patients with meta-static colorectal cancer. BMJ. 306:752–755. 1993. View Article : Google Scholar : PubMed/NCBI
11	Mimeault M, Hauke R and Batra SK: Recent advances on the molecular mechanisms involved in the drug resistance of cancer cells and novel targeting therapies. Clin Pharmacol Ther. 83:673–691. 2008. View Article : Google Scholar
12	Flaten GE, Dhanikula AB, Luthman K and Brandl M: Drug permeability across a phospholipid vesicle based barrier: a novel approach for studying passive diffusion. Eur J Pharm Sci. 27:80–90. 2006. View Article : Google Scholar
13	Goldberg RM, Sargent DJ, Morton RF, Fuchs CS, Ramanathan RK, Williamson SK, Findlay BP, Pitot HC and Alberts SR: A randomized controlled trial of fluorouracil plus leucovorin, irinotecan, and oxaliplatin combinations in patients with previously untreated metastatic colorectal cancer. J Clin Oncol. 22:23–30. 2004. View Article : Google Scholar
14	Dinu-Pîrvu C, Ferdes M, Buţu A, Orţan A and Ghica MV: Physicochemical investigation of low soluble biocompounds entrapped in lipid carriers. Farmacia. 61:182–192. 2013.
15	House MG, Ito H, Gönen M, Fong Y, Allen PJ, DeMatteo RP, Brennan MF, Blumgart LH, Jarnagin WR and D'Angelica MI: Survival after hepatic resection for metastatic colorectal cancer: trends in outcomes for 1,600 patients during two decades at a single institution. J Am Coll Surg. 210:744–752. 752–755. 2010. View Article : Google Scholar : PubMed/NCBI
16	Abbott A: Cell culture: Biology's new dimension. Nature. 424:870–872. 2003. View Article : Google Scholar : PubMed/NCBI
17	Mazzoleni G, Di Lorenzo D and Steimberg N: Modelling tissues in 3D: The next future of pharmaco-toxicology and food research? Genes Nutr. 4:13–22. 2009. View Article : Google Scholar :
18	Pampaloni F, Reynaud EG and Stelzer EH: The third dimension bridges the gap between cell culture and live tissue. Nat Rev Mol Cell Biol. 8:839–845. 2007. View Article : Google Scholar : PubMed/NCBI
19	Ghosh S, Spagnoli GC, Martin I, Ploegert S, Demougin P, Heberer M and Reschner A: Three-dimensional culture of melanoma cells profoundly affects gene expression profile: A high density oligonucleotide array study. J Cell Physiol. 204:522–531. 2005. View Article : Google Scholar : PubMed/NCBI
20	Ernst A, Hofmann S, Ahmadi R, Becker N, Korshunov A, Engel F, Hartmann C, Felsberg J, Sabel M, Peterziel H, et al: Genomic and expression profiling of glioblastoma stem cell-like spheroid cultures identifies novel tumor-relevant genes associated with survival. Clin Cancer Res. 15:6541–6550. 2009. View Article : Google Scholar : PubMed/NCBI
21	Haycock JW: 3D cell culture: A review of current approaches and techniques. Methods Mol Biol. 695:1–15. 2011. View Article : Google Scholar
22	Sutherland RM, Sordat B, Bamat J, Gabbert H, Bourrat B and Mueller-Klieser W: Oxygenation and differentiation in multicellular spheroids of human colon carcinoma. Cancer Res. 46:5320–5329. 1986.PubMed/NCBI
23	Hirschhaeuser F, Menne H, Dittfeld C, West J, Mueller-Klieser W and Kunz-Schughart LA: Multicellular tumor spheroids: An underestimated tool is catching up again. J Biotechnol. 148:3–15. 2010. View Article : Google Scholar : PubMed/NCBI
24	Spencer VA, Xu R and Bissell MJ: Gene expression in the third dimension: The ECM-nucleus connection. J Mammary Gland Biol Neoplasia. 15:65–71. 2010. View Article : Google Scholar : PubMed/NCBI
25	Jacks T and Weinberg RA: Taking the study of cancer cell survival to a new dimension. Cell. 111:923–925. 2002. View Article : Google Scholar
26	Sutherland RM, McCredie JA and Inch WR: Growth of multicell spheroids in tissue culture as a model of nodular carcinomas. J Natl Cancer Inst. 46:113–120. 1971.PubMed/NCBI
27	Singh SK, Clarke ID, Terasaki M, Bonn VE, Hawkins C, Squire J and Dirks PB: Identification of a cancer stem cell in human brain tumors. Cancer Res. 63:5821–5828. 2003.PubMed/NCBI
28	Deisboeck TS, Berens ME, Kansal AR, Torquato S, Stemmer-Rachamimov AO and Chiocca EA: Pattern of self-organization in tumour systems: Complex growth dynamics in a novel brain tumour spheroid model. Cell Prolif. 34:115–134. 2001. View Article : Google Scholar : PubMed/NCBI
29	Wartenberg M, Dönmez F, Ling FC, Acker H, Hescheler J and Sauer H: Tumor-induced angiogenesis studied in confrontation cultures of multicellular tumor spheroids and embryoid bodies grown from pluripotent embryonic stem cells. FASEB J. 15:995–1005. 2001. View Article : Google Scholar : PubMed/NCBI
30	Xiang X, Phung Y, Feng M, Nagashima K, Zhang J, Broaddus VC, Hassan R, Fitzgerald D and Ho M: The development and characterization of a human mesothelioma in vitro 3D model to investigate immunotoxin therapy. PLoS One. 6:e146402011. View Article : Google Scholar : PubMed/NCBI
31	Del Duca D, Werbowetski T and Del Maestro RF: Spheroid preparation from hanging drops: Characterization of a model of brain tumor invasion. J Neurooncol. 67:295–303. 2004. View Article : Google Scholar : PubMed/NCBI
32	Friedrich J, Seidel C, Ebner R and Kunz-Schughart LA: Spheroid-based drug screen: Considerations and practical approach. Nat Protoc. 4:309–324. 2009. View Article : Google Scholar : PubMed/NCBI
33	Weiswald LB, Guinebretière JM, Richon S, Bellet D, Saubaméa B and Dangles-Marie V: In situ protein expression in tumour spheres: Development of an immunostaining protocol for confocal microscopy. BMC Cancer. 10:1062010. View Article : Google Scholar : PubMed/NCBI
34	Li Q, Chen C, Kapadia A, Zhou Q, Harper MK, Schaack J and LaBarbera DV: 3D models of epithelial-mesenchymal transition in breast cancer metastasis: High-throughput screening assay development, validation, and pilot screen. J Biomol Screen. 16:141–154. 2011. View Article : Google Scholar : PubMed/NCBI
35	Wright MH, Calcagno AM, Salcido CD, Carlson MD, Ambudkar SV and Varticovski L: Brca1 breast tumors contain distinct CD44+/CD24− and CD133+ cells with cancer stem cell characteristics. Breast Cancer Res. 10:R102008. View Article : Google Scholar
36	Takaishi S, Okumura T, Tu S, Wang SS, Shibata W, Vigneshwaran R, Gordon SA, Shimada Y and Wang TC: Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells. 27:1006–1020. 2009. View Article : Google Scholar : PubMed/NCBI
37	Lee GY, Kenny PA, Lee EH and Bissell MJ: Three-dimensional culture models of normal and malignant breast epithelial cells. Nat Methods. 4:359–365. 2007. View Article : Google Scholar : PubMed/NCBI
38	Fischbach C, Chen R, Matsumoto T, Schmelzle T, Brugge JS, Polverini PJ and Mooney DJ: Engineering tumors with 3D scaffolds. Nat Methods. 4:855–860. 2007. View Article : Google Scholar : PubMed/NCBI
39	Kim JB, Stein R and O'Hare MJ: Three-dimensional in vitro tissue culture models of breast cancer - a review. Breast Cancer Res Treat. 85:281–291. 2004. View Article : Google Scholar : PubMed/NCBI
40	Derycke AS and De Witte PA: Transferrin-mediated targeting of hypericin embedded in sterically stabilized PEG-liposomes. Int J Oncol. 20:181–187. 2002.
41	Ion RM and Apostol S: Photochemical study of porphyrin aggregates. Rev Chem. 56:607–620. 2005.
42	Mueller-Klieser W: Multicellular spheroids: A review on cellular aggregates in cancer research. J Cancer Res Clin Oncol. 113:101–122. 1987. View Article : Google Scholar
43	Wibe E, Berg JP, Tveit KM, Nesland JM and Lunde S: Multicellular spheroids grown directly from human tumour material. Int J Cancer. 34:21–26. 1984. View Article : Google Scholar : PubMed/NCBI
44	Boothby CD, Daniel J, Adam S and Dyson JE: Use of a laser diffraction particle sizer for the measurement of mean diameter of multicellular tumour spheroids. In Vitro Cell Dev Biology. 25:946–950. 1989. View Article : Google Scholar
45	Olive PL and Durand RE: Drug and radiation resistance in spheroids: cell contact and kinetics. Cancer Metastasis Rev. 13:121–138. 1994. View Article : Google Scholar : PubMed/NCBI
46	Inoue T, Inoue T, Teshima T, Murayama S, Shimizutani K, Fuchihata H and Furukawa S: Phase III trial of high and low dose rate interstitial radiotherapy for early oral tongue cancer. Int J Radiat Oncol Biol Phys. 36:1201–1204. 1996. View Article : Google Scholar : PubMed/NCBI