Research Paper Volume 13, Issue 4 pp 5284—5296

Matrix stiffness promotes glioma cell stemness by activating BCL9L/Wnt/β-catenin signaling

Wnt/β-catenin signaling was activated in glioma cells cultured on high-stiffness gels. (A) Western blotting analysis of β-catenin expression in LN229 cells cultured on gels of different stiffness levels with or without BCL9L shRNA treatment. (B) Western blotting analysis of β-catenin expression in T98G cells cultured on gels of different stiffness levels with or without BCL9L shRNA treatment. (C) Flow cytometry analysis of CD133 expression in LN229 or T98G cells cultured on 16-kPa stiffness gels and treated with PBS, gigantol (100 μM, 72 h) or KYA1797K (25 μM, 72 h). (D) Proliferation of LN229 or T98G cells treated with PBS, gigantol (100 μM, 72 h) or KYA1797K (25 μM, 72 h) and cultured on 16-kPa stiffness gels. (E) Tumor volumes were measured at various time points in mice injected with LN229 or T98G cells that had been treated with PBS, gigantol (100 μM, 72 h) or KYA1797K (25 μM, 72 h) and cultured on 16-kPa stiffness gels. (F) Cell colony formation of LN229 or T98G cells treated with PBS, gigantol (100 μM, 72 h) or KYA1797K (25 μM, 72 h) and cultured on 16-kPa stiffness gels. (G) Tumorigenesis of mice subcutaneously injected with 104 LN229 or T98G cells that had been pre-treated with PBS, gigantol (100 μM, 72 h) or KYA1797K (25 μM, 72 h) and cultured on 16-kPa stiffness gels. (H) Immunofluorescence analysis of β-catenin expression in tissues derived from the HS and LS groups. Scale bar, 50 μm. *P

Figure 3. Wnt/β-catenin signaling was activated in glioma cells cultured on high-stiffness gels. (A) Western blotting analysis of β-catenin expression in LN229 cells cultured on gels of different stiffness levels with or without BCL9L shRNA treatment. (B) Western blotting analysis of β-catenin expression in T98G cells cultured on gels of different stiffness levels with or without BCL9L shRNA treatment. (C) Flow cytometry analysis of CD133 expression in LN229 or T98G cells cultured on 16-kPa stiffness gels and treated with PBS, gigantol (100 μM, 72 h) or KYA1797K (25 μM, 72 h). (D) Proliferation of LN229 or T98G cells treated with PBS, gigantol (100 μM, 72 h) or KYA1797K (25 μM, 72 h) and cultured on 16-kPa stiffness gels. (E) Tumor volumes were measured at various time points in mice injected with LN229 or T98G cells that had been treated with PBS, gigantol (100 μM, 72 h) or KYA1797K (25 μM, 72 h) and cultured on 16-kPa stiffness gels. (F) Cell colony formation of LN229 or T98G cells treated with PBS, gigantol (100 μM, 72 h) or KYA1797K (25 μM, 72 h) and cultured on 16-kPa stiffness gels. (G) Tumorigenesis of mice subcutaneously injected with 104 LN229 or T98G cells that had been pre-treated with PBS, gigantol (100 μM, 72 h) or KYA1797K (25 μM, 72 h) and cultured on 16-kPa stiffness gels. (H) Immunofluorescence analysis of β-catenin expression in tissues derived from the HS and LS groups. Scale bar, 50 μm. *P < 0.05, **P < 0.01, n.s. no significant difference.