Glioma stem cells enhance endothelial cell migration and proliferation via the Hedgehog pathway

Yan,Guang‑Ning; Lv,Yang-Fan; Yang,Lang; Yao,Xiao‑Hong; Cui,You‑Hong; Guo,De‑Yu

doi:10.3892/ol.2013.1569

Glioma stem cells enhance endothelial cell migration and proliferation via the Hedgehog pathway

Authors:
- Guang‑Ning Yan
- Yang-Fan Lv
- Lang Yang
- Xiao‑Hong Yao
- You‑Hong Cui
- De‑Yu Guo
View Affiliations

Affiliations: Department of Pathology, Southwest Hospital, Third Military Medical University of PLA, Shapingba District, Chongqing 400038, P.R. China
Published online on: September 9, 2013 https://doi.org/10.3892/ol.2013.1569
Pages: 1524-1530

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 aim of the present study was to determine the possible mechanism underlying the enhanced migration and proliferation of endothelial cells caused by glioma stem cells (GSCs). Tumor spheres enriched in GSCs derived from the mouse GL261 glioma cell line, and the brain microvessel endothelial cell line, b.END3, were used in this study. A Transwell co‑culture system, RNAi experiments, quantitative polymerase chain reaction, western blotting and enzyme‑linked immunosorbent, cell counting kit‑8 (CCK‑8) proliferation, Transwell migration and wound‑healing assays were used in this study to determine the migration and proliferation ability, as well as the Hedgehog (HH) pathway‑related gene expression in the b.END3 cells. Based on the results, it was demonstrated that the migration and proliferation of the endothelial cells were enhanced following co‑culture with GSCs. The gene expression of the HH pathway‑related genes, Sonic Hedgehog (Shh) and Hedgehog‑interacting protein (Hhip) was altered in the endothelial cells when co‑cultured with GSCs. Overexpression of glioma‑associated oncogene homolog 1 indicated activation of the HH pathway. Following knockdown of smoothened (Smo) in the endothelial cells, the migration and proliferation abilities of the cells were inhibited. GSCs have little effect on enhancing these behaviors in endothelial cells following Smo‑knockdown. Further investigation revealed that Shh levels in the supernatant of the co‑culture system were elevated, indicating the importance of secreted Shh from the endothelial cells. In conclusion, GSCs enhanced the migration and proliferation of the endothelial cells in vitro, which was likely associated with the activation of the HH pathway in the endothelial cells, caused by the increased secretion of Shh.

View Figures

View References

1	Berger MS: Glioma surgery: a century of challenge. Clin Neurosurg. 58:7–9. 2011. View Article : Google Scholar : PubMed/NCBI
2	Quick A, Patel D, Hadziahmetovic M, et al: Current therapeutic paradigms in glioblastoma. Rev Recent Clin Trials. 5:14–27. 2010. View Article : Google Scholar : PubMed/NCBI
3	Chaichana KL, McGirt MJ, Laterra J, et al: Recurrence and malignant degeneration after resection of adult hemispheric low grade gliomas. J Neurosurg. 112:10–17. 2010. View Article : Google Scholar : PubMed/NCBI
4	Dietrich J, Diamond EL and Kesari S: Glioma stem cell signaling: therapeutic opportunities and challenges. Expert Rev Anticancer Ther. 10:709–722. 2010. View Article : Google Scholar : PubMed/NCBI
5	O'Brien CA, Kreso A and Jamieson CH: Cancer stem cells and self-renewal. Clin Cancer Res. 16:3113–3120. 2010. View Article : Google Scholar
6	Liu Y, Carson-Walter EB, Cooper A, et al: Vascular gene expression patterns are conserved in primary and metastatic brain tumors. J Neurooncol. 99:13–24. 2010. View Article : Google Scholar : PubMed/NCBI
7	Calabrese C, Poppleton H, Kocak M, et al: A perivascular niche for brain tumor stem cells. Cancer Cell. 11:69–82. 2007. View Article : Google Scholar : PubMed/NCBI
8	Li M, Niu C, Gao G, et al: The study of the relation between proliferating brain tumor stem cells with micro-vascular system. Chin J Neurooncol. 8:82–87. 2010.(In Chinese).
9	Barami K: Relationship of neural stem cells with their vascular niche: implications in the malignant progression of gliomas. J Clin Neurosci. 15:1193–1197. 2008. View Article : Google Scholar : PubMed/NCBI
10	Fu JR, Liu WL, Zhou JF, et al: Sonic hedgehog protein promotes bone marrow-derived endothelial progenitor cell proliferation, migration and VEGF production via PI 3-kinase/Akt signaling pathways. Acta Pharmacol Sin. 27:685–693. 2006. View Article : Google Scholar : PubMed/NCBI
11	Huang Q, Dong J, Zhu Y, et al: Isolation and culture of tumor stem cells from human brain glioma tissues. Zhonghua Zhong Liu Za Zhi. 28:331–333. 2006.(In Chinese).
12	Zhou K, Yang H, Zhou W, Zhang C, Shu H and Wang B: Effect of drug blockade of type-3 metabotropic glutamate receptor on proliferation and differentiation in glioma stem cells. Di 3 Jun Yi Da Xue Xue Bao. 34:1701–1706. 2012.(In Chinese).
13	Hess KR, Broglio KR and Bondy ML: Adult glioma incidence trends in the United States, 1977–2000. Cancer. 101:2293–2299. 2004.PubMed/NCBI
14	Singh SK, Clarke ID, Terasaki M, et al: Identification of a cancer stem cell in human brain tumors. Cancer Res. 63:5821–5828. 2003.PubMed/NCBI
15	Charalambous C, Chen TC and Hofman FM: Characteristics of tumor-associated endothelial cells derived from glioblastoma multiforme. Neurosurg Focus. 20:E222006. View Article : Google Scholar : PubMed/NCBI
16	Bao S, Wu Q, Sathornsumetee S, et al: Stem cell-like glioma cells promote tumor angiogenesis through vascular endothelial growth factor. Cancer Res. 66:7843–7848. 2006. View Article : Google Scholar : PubMed/NCBI
17	Borovski T, De Sousa E, Melo F, Vermeulen L and Medema JP: Cancer stem cell niche: the place to be. Cancer Res. 71:634–639. 2011. View Article : Google Scholar : PubMed/NCBI
18	Wang R, Chadalavada K, Wilshire J, et al: Glioblastoma stem-like cells give rise to tumour endothelium. Nature. 468:829–833. 2010. View Article : Google Scholar : PubMed/NCBI
19	Folkins C, Shaked Y, Man S, et al: Glioma tumor stem-like cells promote tumor angiogenesis and vasculogenesis via vascular endothelial growth factor and stromal-derived factor 1. Cancer Res. 69:7243–7251. 2009. View Article : Google Scholar : PubMed/NCBI
20	Scharpfenecker M, van Dinther M, Liu Z, et al: BMP-9 signals via ALK1 and inhibits bFGF-induced endothelial cell proliferation and VEGF-stimulated angiogenesis. J Cell Sci. 120:964–972. 2007. View Article : Google Scholar : PubMed/NCBI
21	Yao XH, Ping YF, Chen JH, et al: Glioblastoma stem cells produce vascular endothelial growth factor by activation of a G-protein coupled formylpeptide receptor FPR. J Pathol. 215:369–376. 2008. View Article : Google Scholar
22	Ping YF, Yao XH, Jiang JY, et al: The chemokine CXCL12 and its receptor CXCR4 promote glioma stem cell-mediated VEGF production and tumor angiogenesis via PI3K/AKT signalling. J Pathol. 224:344–354. 2011. View Article : Google Scholar : PubMed/NCBI
23	Medici D and Olsen BR: Rapamycin inhibits proliferation of hemangioma endothelial cells by reducing HIF-1-dependent expression of VEGF. PLoS One. 7:e429132012. View Article : Google Scholar : PubMed/NCBI
24	Ma G, Xiao Y and He L: Recent progress in the study of Hedgehog signaling. J Genet Genomics. 35:129–137. 2008. View Article : Google Scholar : PubMed/NCBI
25	Asai J, Takenaka H, Kusano KF, et al: Topical sonic hedgehog gene therapy accelerates wound healing in diabetes by enhancing endothelial progenitor cell-mediated microvascular remodeling. Circulation. 113:2413–2424. 2006. View Article : Google Scholar