Arsenic trioxide inhibits Ewing’s sarcoma cell invasiveness by targeting p38^MAPK and c-Jun N-terminal kinase

Abstract

Ewing’s sarcoma is the second most frequent primary malignant bone tumor, mainly affecting children and young adults. The notorious metastatic capability of this tumor aggravates patient mortality and remains a problem to be overcome. We investigated the effect of arsenic trioxide (As₂O₃) on the metastasis capability of Ewing’s sarcoma cells. We performed 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide assays to choose appropriate concentrations of As₂O₃ for the experiments. Migration, invasion, and adhesion assays were performed to assess the effect of As₂O₃ on the metastasis of Ewing’s sarcoma. Immunofluorescent staining was used to observe cytoskeleton reorganization in Ewing’s sarcoma cells treated with As₂O₃. Changes in matrix metalloproteinase-9 expression and the mitogen-activated protein kinase (MAPK) pathway were investigated using western blot. Inhibitors of p38^MAPK (sb202190) and c-Jun NH₂-terminal kinase (JNK, sp600125) were used in invasion assays to determine the effect of p38^MAPK and JNK. We found that As₂O₃ may markedly inhibit the migration and invasion capacity of Ewing’s sarcoma cells with structural rearrangements of the actin cytoskeleton. The expressions of matrix metalloproteinase-9, phosphor-p38^MAPK, and phosphor-JNK were suppressed by As₂O₃ treatment in a dose-dependent manner. The inhibitors of p38^MAPK (sb202190) and JNK (sp600125) enhanced the inhibition induced by As₂O_3, which was counteracted by anisomycin, an activating agent of p38^MAPK and JNK. Taken together, our results demonstrate that As₂O₃ can inhibit the metastasis capability of RD-ES and A-673 cells and may have new therapeutic value for Ewing’s sarcoma.