Aim: The study aims to evaluate the in-vitro anti-cancer effect of the ethanolic extract of Xanthium indicum Koenig (Asteraceae) Materials and Methods: The plant was chosen and extracted by Maceration method. The extraction was processed using the following solvents such as Petroleum ether, Chloroform, ethyl acetate, and ethanol. TLC and HPTLC analysis was performed using the solvents, and the Rf values was noted. The in vitro antioxidant using DPPH scavenging assay and in-vitro anti-cancer activity using MTT assay and Apoptosis by ethanolic extract of Xanthium indicum plant. The IC50 value was noted accordingly. Result: Ethanol was considered as a better solvent; in each test, only ethanol showed comparatively high value than others. MTT assay revealed that the viability of the cell increases with increase in concentration, similarly in Apoptosis method cell death was seen as concentration increases. Conclusion: Ethanolic extract of Xanthium Indicum was found to possess high potentiality. Hence, it can be subjected to study further, and prominent anti-cancer research can be done.