Glioblastoma multiforme (GBM), the most malignant gliomas, has remarkable tendency to infiltrate through surrounding normal brain tissue, which confers resistance to most of current therapies. Previously reported histologic evidences have shown that glioma cells migrate along the orientation of thin, elongated anatomic structures such as white matter tract and capillary vessel walls. Understanding topographical effects on glioma cell migration is important to develop new anti-migratory therapies; however, conventional culture systems have limitations to reproduce native environment for cell migration.In this study, we tried to identify the topographical effect on cell migration and its phenotype by culturing glioma cells on micro-patterned substrates mimicking brain anatomy. Glioma cells on micro-patterned substrates showed elongated morphology, highly aligned pattern and increased migration along the patterned direction. In contrast, the glioma cells on non-patterned substrates showed round morphology and random orientation with less migratory activity. Accordingly, the molecular feature of glioma cells was shown differently depending on surface topography, suggesting the impacts on intracellular signal by mechanical cues. In order to figure out the mechanism of phenotype change by mechanical cues, we examined the inhibition effects of associated signaling pathways on glioma cells. Glioma cells on flat and grooved substrates responded in different manner, which might be dependent on topography of substrates. Consequently, integrin might be considered as a mechanosensitive molecule of topographical cues and mediator for mechanotransduction.In conclusion, glioma cell migration and its intracellular signaling are highly regulated by mechanical cues mimicking neural topography. Thus, understanding mechanical effects on cells using micro-patterned scaffolds can contribute to identify effective anti-migratory therapies.