SALINOMYCIN PREVENTS ANGIOGENESIS AND INHIBITS GLIOMA GROWTH BY INTERFERING WITH VEGF-VEGFR2-AKT/FAK SIGNAL AXIS

Wenqianjun Sheng1, Ruofei Jia1,#,  Jun Wang**, Jun Xia**

*Foshan Hospital of TCM, Foshan,PR China - **The first people’s hospital of Changde city, Changde, PR China

Objective: To analyse the mechanism of salinomycin preventing angiogenesis and inhibiting glioma growth by interfering with VEGF-VEGFR2-AKT/FAK signal axis.

Methods: Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and treated with different concentrations of salinomycin (0, 1, 2, 4, 8 μM) for 48 h, with positive control (vascular endothelial growth factor (VEGF)) and negative control (DDP) set up. The cell morphology was observed under an inverted microscope, the cell activity was detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the invasion ability and migration ability of HUVECs after salinomycin treatment were evaluated by in vitro invasion experiment and scratch test, and the vascular formation ability of HUVECs treated with salinomycin was evaluated by angiogenesis assay. The effect of salinomycin on the expression of VEGF receptor and other pathway proteins was detected by Western blot assay. The U251 glioma cell model was established and the control group was created. The tumour specimens were analysed by Western blot assay and immunohistochemistry method.

Results: The results of MTT assay showed that the effect of different concentrations of salinomycin on the growth of HUVECs was dose-dependent. The inverted microscope showed that the number of cells decreased, while the morphology atrophied and became round. The results of the scratch test, in vitro invasion experiment and the angiogenic assay showed that salinomycin could inhibit the angiogenesis of HUVECs. Western blot assay also showed that the level of P-FAK decreased significantly and the expression of VEGF, p-VEGFR2 and p-AKT decreased after salinomycin treatment, suggesting that salinomycin interferes with VEGF-VEGFR2-AKT/FAK signalling pathway. The volume and weight of the tumour were reduced under the action of salinomycin and the expression of P-FAK and p-AKT in the tissue decreased. 

Conclusion: Salinomycin can block angiogenesis by interfering with VEGF-VEGFR2-AKT/FAK signalling pathway, thereby inhibiting the growth of glioma.