Background: Chlorogenic acid (CA), a polyphenolic component of fruits, vegetables, coffee, wine, and olive oil, has beneficial effects on human heath, including antioxidant and anticancer effects. However, its precise effects on glioma have not been examined. Objective: Our study aimed to explore the anticancer effects of CA obtained from Viscum album callus on C6 glioma cell migration and proliferation. Materials and Methods: Anticancer potency was analyzed by the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt assay to assess the ability to inhibit cell growth and proliferation. Cell mobility was investigated based on the Boyden chamber and the scratch wound healing assay. Factors involved in cell cycle progression were evaluated by mRNA and protein expression. Cell death was determined by staining with specific dyes and fluorescence microscopy. Results: CA significantly reduced C6 glioma cell proliferation and migration. Furthermore, it induced reactive oxygen species generation and apoptotic cell death. Treatment with CA also suppressed extracellular signal-regulated kinase ½ (ERK½) phosphorylation and the gene expression of cyclins E and A. Conclusion: Our results show that CA may regulate glioma cell migration and proliferation via modulation of ERK½ phosphorylation and cell cycle regulation. Thus, it might be a potent anticancer agent in preventing progression of glioma.