Colorectal cancer (CRC) recorded to be the second most deadly cancer in world, treatment of these type of cancers have severe chemotherapeutic side effects. Alternative medicine comes from the folk knowledge of indigenous people and these plant-derived drugs were originally explored through the study of traditional cures with very minimal side effects. Flowers of Musa acuminata possess a wide range of pharmacological properties such as antihypertensive, immunomodulatory, anti HIV, anti-tumor activity, Anti-Helmintic, hence the present study was aimed to evaluate effect of Musa acuminata flower extract in Human colon cancer cell line Colo320.The primary phytochemical analysis of methanolic extract of Musa acuminata flower exposed the presence various phytochemical such as alkaloids, flavonoids, steroids, tannins, total phenols. The assays of enzymatic antioxidants in the methanolic extract of Musa acuminata flower shows a notable amount of Super oxide dismutase (14U/g) in it. methanolic extract of Musa acuminata flower contains good amount of vitamin C, vitamin A and E. The methanolic extract of Musa acuminata flower showed the evidence of induction of apoptosis in the colo320 cell line, were confirmed by DNA fragmentation studies and AO/EB staining. Thus the methanolic extract of Musa acuminata flower has an inhibitory role on human colon cancer cell line Colo320.