Figure 2

Inhibition of paediatric glioma tumour sphere invasion by LiCl and BIO. Tumour spheroids (SF188) were embedded in collagen and incubated in (A) LiCl at 40, 20, 10, 5 mM in NaCl or (B) BIO at 10, 5, 1, 0.5 μ M in DMSO for 72 h. Migration was inhibited by both LiCl and BIO as shown in representative stills at 24, 48 and 72 h using the EVOS cell imaging system at × 40 magnification. Scale bar=1000 μm. (C) Quantitative analysis of migration inhibition confirmed the results visualised by microscopy. Control medium used was Dulbecco’s Modified Eagles’ medium plus 10% heat-inactivated foetal calf serum. Both LiCl and BIO had statistically significant anti-migratory effects on the invasion zone and leading edge as indicated by * on all three cell lines (SF188, KNS42 and HSJD-DIPG-007). Results are representative of n=3 individual experiments. Error bars expressed as ±s.e.m. *P<0.05, **P<0.01, ***P<0.001 by ANOVA for each cell type and treatment. (D) Cell viability of paediatric glioma lines (SF188 and KNS42) grown as tumour spheroids treated with various concentrations of LiCl and BIO was determined by WST-1 assay and then expressed as a percentage of controls. Error bars expressed as ±s.e.m. The effects of LiCl and BIO on β-catenin localisation were evaluated for SF188 (E) and KNS42 (F) compared with medium mock-treated controls by immunofluorescent labelling. Arrows indicate β-catenin localisation. Red labelling–β-catenin, green labeling–actin, blue labeling–DAPI staining. Magnification × 63.