Isoflurane enhances the malignant potential of glioblastoma stem cells by promoting their viability, mobility in vitro and migratory capacity in vivo

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Isoflurane enhances the malignant potential of glioblastoma stem cells by promoting their viability, mobility in vitro and migratory capacity in vivo M Zhu, M Li, Y Zhou, S Dangelmajer, U.D. Kahlert, R Xie, Q Xi, A Shahveranov, D Ye, T Lei British Journal of Anaesthesia Volume 116, Issue 6, Pages 870-877 (June 2016) DOI: 10.1093/bja/aew124 Copyright © 2016 The Author(s) Terms and Conditions

Fig 1 Proliferation capacity of U251-GSCs in vitro. (a) Representative image of Ki-67 (pink) and DAPI (blue) staining and (b) quantitative analysis of staining in U251-GSCs to isoflurane for 3 h. (c) Representative image of Ki-67 (pink) and DAPI (blue) staining and (d) quantitative analysis of staining in U251-GSCs to isoflurane for 6 h. Scale bars+50 μm. Mean (sd) shown; n+9 per group. *P<0.05, **P<0.01, ***P<0.001. British Journal of Anaesthesia 2016 116, 870-877DOI: (10.1093/bja/aew124) Copyright © 2016 The Author(s) Terms and Conditions

Fig 2 Apoptosis of U251-GSCs in vitro. (a) Representative image of flow cytometry analysis of annexin V and propidium iodide as a measure of apoptosis and (b) quantitative analysis of apoptosis in U251-GSCs exposed to isoflurane for 3 h. (c) Representative image of flow cytometry analysis of annexin V and propidium iodide as a measure of apoptosis and (d) quantitative analysis of apoptosis in U251-GSCs exposed to isoflurane for 6 h. Mean (sd) shown; n+3 per group. *P<0.05, **P<0.01, ***P<0.001. British Journal of Anaesthesia 2016 116, 870-877DOI: (10.1093/bja/aew124) Copyright © 2016 The Author(s) Terms and Conditions

Fig 3 Migration capacity of U251-GSCs in vitro. (a) Representative image of transwell analysis of migration and (b) quantitative analysis of migration of U251-GSCs exposed to isoflurane for 3 h. (c) Representative image of transwell analysis of migration and (d) quantitative analysis of migration of U251-GSCs exposed to isoflurane for 6 h. Mean (SD) shown; n+9 per group. *P<0.05, **P<0.01, ***P<0.001. British Journal of Anaesthesia 2016 116, 870-877DOI: (10.1093/bja/aew124) Copyright © 2016 The Author(s) Terms and Conditions

Fig 4 Tumour migratory potential capacity and tumour volume in vivo. (a) Representative pictures of DAPI (blue) and human nuclei (green) staining in mice brain after injection of GSCs pretreated with isoflurane. (b) Schematic diagram showing quantification of tumour volume and (c) quantitative analysis of tumour volume analysis in mouse brain. (d) Schematic diagram showing quantification of migration distance and (e) quantitative analysis of tumour volume analysis in mouse brain. Scale bars+100 μm. Mean (sd) shown; n+15 per group. *P<0.05, **P<0.01, ***P<0.001. British Journal of Anaesthesia 2016 116, 870-877DOI: (10.1093/bja/aew124) Copyright © 2016 The Author(s) Terms and Conditions