Surface topography was believed to be a factor affecting cellular morphology, proliferation, and differentiation. In present work, we examined the facts that nanogrooved surfaces had an effect on the attachment and alignment behavior of C6 cells in detail. The nanogrooves on the surface were fabricated by Laser-induced periodic structures (LIPS) technique (Polarized laser:λ=266nm). The parallel grooves and ridges on the polystyrene were 300-350nm in period and 40-50nm in depth. In addition, we also tried to control cells growth via surface nanostructures on the patterns. And then we evaluated the attachment and alignment of C6 cells on these substrates. After 2 days of incubation, we found that C6 cells were elongated and aligned along patterns of grooves and ridges, whereas were mostly random on smooth substrates. By observing, the response of cells to nano-sized feature suggested that the formation of fibrous cellular components, especially in the filopodium and lamellipodia, were influenced by the nanogrooves. These findings would have potential applications in tissue engineering and in the design of biosensors.