Abstract Objectives: Medulloblastoma is the most common malignant and metastatic pediatric brain tumor in children. Angiogenesis inhibitors, such as sunitinib, represent a promising strategy to improve glioblastoma (GBM) tumor response. Temozolomide (TMZ) also became the standard of care for GBM patients by removing alkyl groups from the O6-position of guanine. But, there is little evidence about migration and invasion study on medulloblastoma cells treated with sunitinib or TMZ. In this study, we identified the effects of sunitinib or TMZ only, and the combination of them on migration and invasiveness in medulloblastoma cell line (DAOY). Also the characteristics of cancer stem cells were investigated. Methods and Results: The cell proliferation rate showed TMZ (1000µM) 70%, sunitinib (62.5µM) 60%, and the combination of TMZ and sunitinib (1000/62.5µM) 35% at 72hr after treatment with the drugs in DAOY cells. Treatment with sunitinib in addition to TMZ in the cells demonstrated more G2/M arrest than cells treated respective drugs at 24hr exposure of the drugs from cell cycle analysis. Through Rhodamine phalloidin staining, we also observed change of cell size and F-actin alterations in all cells treated TMZ, sunitinib only or the combination of the reagents, in contrast to cells treated DMSO as a control. And, in invasion assay using Matrigel chamber, the cells treated with combination of drugs were inhibited invasiveness the most when compared with control, sunitinib, and TMZ only at 72hr after exposure to the drugs. Moreover, when the cells were treated with sunitinib and TMZ simultaneously, migration ability from wound-healing assay represented higher than in cells treated TMZ or sunitinib only during exposure of 24hr. We investigated properties of cancer stem cells as well as characteristics of migration and invasion in cells treated the drugs analyzing the expression of protein during exposure of 24, 48, 72hr by western blotting. As a result, we identified the decreased levels of the protein such as p-PDGFR-β, p-VEGFR1, integrin-β1, MMP-9, CD44, Rac1 as migration and invasion markers, nestin, CD133, Oct3/4, Sox2 as cancer stem cell markers and O6-methylguanine methyltransferase(MGMT). Conclusions: Taken together, the combination therapy functions as very effective therapeutic tool in terms of, not only inhibititor of proliferation, migration and invasiveness, but also cancer stem cells in medulloblastoma cell line. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3321. doi:10.1158/1538-7445.AM2011-3321