Figure 2.

CYY292 reduces GBM cell migration and invasion and suppresses EMT. (A, D) Effect of CYY292, AZD4547, and PD173074 on the viability of GBM cancer cells. Cells were challenged with increasing concentrations of CYY292 for 24 h and cell viability was measured by CCK8. IC₅₀ values in U87MG and LN229 are shown. (B, E) Boyden chamber migration and invasion assays of U87MG (B) and LN229 (E) cells. Cells pretreated with CYY292 at different doses for 24 h were seeded in the upper insert in the presence (for invasion assay) or absence (for migration assay) of pre-coated Matrigel (n = 3). (C, F) Quantification of the migrated and invaded U87MG cells as shown in (B) and the migrated and invaded LN229 cells as shown in (E). (G, H) Immunofluorescent staining of F-actin and cortactin in vehicle- and CYY292-treated LN229 (G) and U87MG (H) cells. Nuclear, DAPI (blue). (I, J) qPCR analysis validated ECM remodeling genes (MMP2, MMP9, MMP13, MMP14), cell growth-stimulating genes (ID1 and ID3) as well as genes encoding EMT (E-cadherin, vimentin, FN1, Krt8). (K) Cell apoptosis analysis of U87MG cells that were treated with vehicle or different doses of CYY292 for 24 h (n = 3). Cells were co-stained with Annexin V and PI. Data are represented as mean ± SD. ∗P < 0.05, ∗∗P < 0.01, one-way ANOVA test.