Abstract Paris polyphylla Smith var. yunnanensis, has been used in traditional Chinese medicine cancer in particular. In this current study, -ecdysterone (1) and three pennogenin steroidal saponins (2-4) were isolated from the EtOH extract of Paris polyphylla var. yunnanensis, and their structures. Their antitumor properties were determined by using an MTT assay in addition to ethidium bromide and acridine orange staining techniques. -1 , respectively. The analysis of the bioactivity-structure relationship shows that the sugar moiety plays a critical role in the activity of steroid moiety. Our results suggest that these three pennogenin steroidal saponins could be utilized to develop anticancer medicines