Background: Emerging drugs aim at targeting the genomic integrity and replication machinery in ovarian cancer. Whereas the antiproliferative activity of Xanthium strumarium L extract (XFC) was recently attributed to alterations in the mitotic apparatus of Chinese hamster ovary (CHO) epithelial cells. XFC capacity to target ovarian cancer cells is unknown. Purpose: Here, we first tested whether XFC impacted on human breast (MCF-7, MDA-MB-231), colorectal (HT-29, C2BBe1) and ovarian cancer cells proliferation, then specifically compared its efficacy in chemosensitive (ES-2) and in cytotoxic drug chemoresistance (SKOV-3) phenotypes of ovarian cancers. Methods: Cell proliferation, survival and, cell cycle assays were used to assess XFC anticancer efficacy. Confocal fluorescent microscopy was used to monitor microtubule alterations, and scratch assays to measure cell migration. Protein expression was quantified by immunobloting. Results: We found that the best proliferation inhibitory effect of XFC was against ovarian cancer cells and ranged from 30-35 µg/ml. XFC efficiently targeted both the cytotoxic drug chemoresistance phenotype of SKOV-3 as well as the chemosensitive ES-2 cells. Cell cycling regulation was found trapped by XFC extract in the G2/M phase in both ES-2 and SKOV-3 cell models. Such effect was, in part, attributable to increased dose-dependent tubulin polymerization which was found higher in SKOV-3 cells. Interestingly, cell cycling regulators Cdk4, Cyclin D3, and p27 were all found decreased in SKOV-3 cells. XFC extracts were effective in inhibiting in vitro migration in both ovarian cancer cell models. Conclusions: Our data support the potential anticancer targeting of chemoresistant human ovarian cancer cells phenotype by XFC extract.