Ilex paraguariensis St. Hilaire tree is native to South America and its dried leaves are used to prepare a traditional beverage called Yerba Mate tea. The aim of this study was to assess the anticancer properties of yerba mate saponins in vitro models. HT-29 (p53 mutant) and RKO (wild type p53) cells were treated with mate saponins (1- 200 μM). Mate saponins inhibited HT-29 (IC50 = 202 μM) and RKO (IC50 = 181 μM) cell proliferation. Mate saponins arrested HT-29 and RKO cells at G1 to S phase by significantly upregulating p21 and p27 proteins, and caused apoptosis through induction of Bax:Bcl-2 protein expression. Mate saponins induced apoptosis and cytotoxicity in human colorectal cancer cells independent of p53 status. Yerba mate tea saponins inhibit human colon cancer cell proliferation.