Three-dimensional hydrogel is suitable for targeted investigation of amoeboid migration of glioma cells

Huang,Yubao; Tong,Luqing; Yi,Li; Zhang,Chen; Hai,Long; Li,Tao; Yu,Shengping; Wang,Wei; Tao,Zhennan; Ma,Haiwen; Liu,Peidong; Xie,Yang; Yang,Xuejun

doi:10.3892/mmr.2017.7888

Three-dimensional hydrogel is suitable for targeted investigation of amoeboid migration of glioma cells

Authors:
- Yubao Huang
- Luqing Tong
- Li Yi
- Chen Zhang
- Long Hai
- Tao Li
- Shengping Yu
- Wei Wang
- Zhennan Tao
- Haiwen Ma
- Peidong Liu
- Yang Xie
- Xuejun Yang
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Affiliations: Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin 300052, P.R. China
Published online on: October 26, 2017 https://doi.org/10.3892/mmr.2017.7888
Pages: 250-256
Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Glioblastoma (GBM) invasion and migration are key biological behaviors leading to refractoriness to current therapies and infiltration into the non‑tumor brain parenchyma. GBM cell migration is strongly dependent on tumor architecture in vivo, which is absent in traditional two‑dimensional (2D) monolayer culture. The present study applied a three‑dimensional (3D) hydrogel model to rebuild the tumor architecture in vitro. Treatment with NSC23766, a specific inhibitor of Ras‑related C3 botulinum toxin substrate 1 (Rac1), inhibited the mesenchymal invasiveness however triggered the amoeboid motility called mesenchymal‑amoeboid transition (MAT). Notably, NSC23766 stimulated U87 GBM cell migration in the 3D hydrogel. However, this compound inhibited cell motility in 2D monolayer culture without tumor architecture for MAT, suggesting the advantage of 3D hydrogel to investigate tumor cell invasion. Due to the inverse interaction of Rac1 and Ras homolog family member A (RhoA) signaling in the transition between mesenchymal and amoeboid morphology, simultaneous treatment of NSC23766 and Y27632 (selective Rho associated coiled‑coil containing protein kinase 1 inhibitor), abolished U87 GBM cell migration through inhibiting MAT and amoeboid‑mesenchymal transition. In addition, Y27632 induced integrin expression which gave rise to the focal adhesion to facilitate the mesenchymal invasion. The results of the present study demonstrated that the 3D hydrogel was a preferable model in vitro to study tumor cell invasion and migration. The combined inhibition of Rac1 and RhoA signaling would be a promising strategy to suppress GBM invasion.

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1	Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJ, Janzer RC, Ludwin SK, Allgeier A, Fisher B, Belanger K, et al: Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol. 10:459–466. 2009. View Article : Google Scholar : PubMed/NCBI
2	Friedl P and Wolf K: Tumour-cell invasion and migration: Diversity and escape mechanisms. Nat Rev Cancer. 3:362–374. 2003. View Article : Google Scholar : PubMed/NCBI
3	Sanz-Moreno V, Gadea G, Ahn J, Paterson H, Marra P, Pinner S, Sahai E and Marshall CJ: Rac activation and inactivation control plasticity of tumor cell movement. Cell. 135:510–523. 2008. View Article : Google Scholar : PubMed/NCBI
4	Rao SS, Bentil S, DeJesus J, Larison J, Hissong A, Dupaix R, Sarkar A and Winter JO: Inherent interfacial mechanical gradients in 3D hydrogels influence tumor cell behaviors. PloS One. 7:e358522012. View Article : Google Scholar : PubMed/NCBI
5	Parri M and Chiarugi P: Rac and Rho GTPases in cancer cell motility control. Cell Commun Signal. 8:232010. View Article : Google Scholar : PubMed/NCBI
6	Jiglaire Jiguet C, Baeza-Kallee N, Denicolaï E, Barets D, Metellus P, Padovani L, Chinot O, Figarella-Branger D and Fernandez C: Ex vivo cultures of glioblastoma in three-dimensional hydrogel maintain the original tumor growth behavior and are suitable for preclinical drug and radiation sensitivity screening. Exp Cell Res. 321:99–108. 2014. View Article : Google Scholar : PubMed/NCBI
7	Zengel P, Nguyen-Hoang A, Schildhammer C, Zantl R, Kahl V and Horn E: µ-Slide chemotaxis: A new chamber for long-term chemotaxis studies. BMC Cell Biol. 12:212011. View Article : Google Scholar : PubMed/NCBI
8	Gouwy M, De Buck M, Pörtner N, Opdenakker G, Proost P, Struyf S and Van Damme J: Serum amyloid A chemoattracts immature dendritic cells and indirectly provokes monocyte chemotaxis by induction of cooperating CC and CXC chemokines. Eur J Immunol. 45:101–112. 2015. View Article : Google Scholar : PubMed/NCBI
9	Thiele J, Ma Y, Bruekers SM, Ma S and Huck WT: 25th anniversary article: Designer hydrogels for cell cultures: A materials selection guide. Adv Mater. 26:125–147. 2014. View Article : Google Scholar : PubMed/NCBI
10	Lu KV, Chang JP, Parachoniak CA, Pandika MM, Aghi MK, Meyronet D, Isachenko N, Fouse SD, Phillips JJ, Cheresh DA, et al: VEGF inhibits tumor cell invasion and mesenchymal transition through a MET/VEGFR2 complex. Cancer Cell. 22:21–35. 2012. View Article : Google Scholar : PubMed/NCBI
11	Mikheeva SA, Mikheev AM, Petit A, Beyer R, Oxford RG, Khorasani L, Maxwell JP, Glackin CA, Wakimoto H, González-Herrero I, et al: TWIST1 promotes invasion through mesenchymal change in human glioblastoma. Mol Cancer. 9:1942010. View Article : Google Scholar : PubMed/NCBI
12	Symons M and Segall JE: Rac and Rho driving tumor invasion: Who's at the wheel? Genome Biol. 10:2132009. View Article : Google Scholar : PubMed/NCBI
13	Yamazaki D, Kurisu S and Takenawa T: Involvement of Rac and Rho signaling in cancer cell motility in 3D substrates. Oncogene. 28:1570–1583. 2009. View Article : Google Scholar : PubMed/NCBI
14	Zhong J, Paul A, Kellie SJ and O'Neill GM: Mesenchymal migration as a therapeutic target in glioblastoma. J Oncol. 2010:4301422010. View Article : Google Scholar : PubMed/NCBI
15	Wuichet K and Søgaard-Andersen L: Evolution and diversity of the Ras superfamily of small GTPases in prokaryotes. Genome Biol Evol. 7:57–70. 2014. View Article : Google Scholar : PubMed/NCBI
16	Haga RB and Ridley AJ: Rho GTPases: Regulation and roles in cancer cell biology. Small GTPases. 7:207–221. 2016. View Article : Google Scholar : PubMed/NCBI
17	Murali A and Rajalingam K: Small Rho GTPases in the control of cell shape and mobility. Cell Mol Life Sci. 71:1703–1721. 2014. View Article : Google Scholar : PubMed/NCBI
18	Rape AD and Kumar S: A composite hydrogel platform for the dissection of tumor cell migration at tissue interfaces. Biomaterials. 35:8846–8853. 2014. View Article : Google Scholar : PubMed/NCBI
19	Ren B, Yu S, Chen C, Wang L, Liu Z, Wu Q, Wang L, Zhao K and Yang X: Invasion and anti-invasion research of glioma cells in an improved model of organotypic brain slice culture. Tumori. 101:390–397. 2015. View Article : Google Scholar : PubMed/NCBI
20	Flemming A: Cancer: Multifunctional nanodevice reverses drug resistance. Nat Rev Drug Discov. 14:3092015. View Article : Google Scholar : PubMed/NCBI
21	Shea LD, Woodruff TK and Shikanov A: Bioengineering the ovarian follicle microenvironment. Annu Rev Biomed Eng. 16:29–52. 2014. View Article : Google Scholar : PubMed/NCBI
22	Souza GR, Molina JR, Raphael RM, Ozawa MG, Stark DJ, Levin CS, Bronk LF, Ananta JS, Mandelin J, Georgescu MM, et al: Three-dimensional tissue culture based on magnetic cell levitation. Nat Nanotechnol. 5:291–296. 2010. View Article : Google Scholar : PubMed/NCBI
23	Reymond N, Im JH, Garg R, Vega FM, d'Agua Borda B, Riou P, Cox S, Valderrama F, Muschel RJ and Ridley AJ: Cdc42 promotes transendothelial migration of cancer cells through β1 integrin. J Cell Biol. 199:653–668. 2012. View Article : Google Scholar : PubMed/NCBI
24	Navarro-Lérida I, Pellinen T, Sanchez SA, Guadamillas MC, Wang Y, Mirtti T, Calvo E and Del Pozo MA: Rac1 nucleocytoplasmic shuttling drives nuclear shape changes and tumor invasion. Dev Cell. 32:318–334. 2015. View Article : Google Scholar : PubMed/NCBI
25	Sadok A, McCarthy A, Caldwell J, Collins I, Garrett MD, Yeo M, Hooper S, Sahai E, Kuemper S, Mardakheh FK and Marshall CJ: Rho kinase inhibitors block melanoma cell migration and inhibit metastasis. Cancer Res. 75:2272–2284. 2015. View Article : Google Scholar : PubMed/NCBI
26	Mishima T, Naotsuka M, Horita Y, Sato M, Ohashi K and Mizuno K: LIM-kinase is critical for the mesenchymal-to-amoeboid cell morphological transition in 3D matrices: Biochem Biophys Res Commun. 392:577–581. 2010.
27	Panková K, Rösel D, Novotný M and Brábek J: The molecular mechanisms of transition between mesenchymal and amoeboid invasiveness in tumor cells. Cell Mol Life Sci. 67:63–71. 2010. View Article : Google Scholar : PubMed/NCBI
28	Desgrosellier JS, Barnes LA, Shields DJ, Huang M, Lau SK, Prévost N, Tarin D, Shattil SJ and Cheresh DA: An integrin alpha(v)beta(3)-c-Src oncogenic unit promotes anchorage-independence and tumor progression. Nat Med. 15:1163–1169. 2009. View Article : Google Scholar : PubMed/NCBI
29	Seguin L, Kato S, Franovic A, Camargo MF, Lesperance J, Elliott KC, Yebra M, Mielgo A, Lowy AM, Husain H, et al: An integrin β₃-KRAS-RalB complex drives tumour stemness and resistance to EGFR inhibition. Nat Cell Biol. 16:457–468. 2014. View Article : Google Scholar : PubMed/NCBI