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Glehnia littoralis (GL) is widely used as an oriental medicine for cough, fever, stroke and other disease conditions. However, the anti‐cancer properties of GL have not been investigated. In order to elucidate the anti‐cancer properties and underlying cell death mechanism, MCF‐7, human breast cancer cells (5 X 104/well) were treated with Glehnia littoralis root extract at 0‐400 ug/ml. The hot water extract of GL root inhibited the proliferation of MCF‐7 cells in both dose‐dependent and time‐dependent manners. Analysis of the cell cycle after treatment of MCF‐7 cells with increasing concentrations of GL root extract for 24 hours showed a significant cell cycle arrest in the G1 phase. RT‐PCR and Western blot analysis both revealed that GL root extract significantly increased the expressions of P21, P27 and P38 while CDK2, CDK4 and Cyclin D1 expressions were down regulated. It is demonstrated that GL root extract arrested the proliferation of MCF‐7 cells at G1 phase through the inhibition of CDK2, CDK4 and Cyclin D1 via an increased induction of P21, P27 and P38. In summary, it is suggested that GL could have possible therapeutic potential against human breast cancer.