Bioassay Guided Fractionation of Allium austroiranicum by Cytotoxic Effects against Ovary and Cervical Cancer Cell Lines

Abstract

Background and objectives: Cancer is a major health problem in the world. The aim of this study was to extract the flowers of Allium austroiranicum used by Iranian people as a condiment or for its medicinal effects followed by bioassay guided fractionation of the extracts and fractions, using anti-proliferative effects against ovarian and cervical cancer cells. Methods: The air-dried flowers of Allium austroiranicum were extracted in a four-step extraction method, resulting hexan, chloroform, chloroform: methanol (9: 1), butanol and aqueous extracts. Anti-proliferative effects of the extracts were evaluated by MTT assay against OVCAR-3, HeLa, and HUVEC cell lines. The most potent cytotoxic extract was then subjected to fractionation by MPLC method on a RP-18 silicagel column. Finally, the cytotoxic effects of resulted fractions were analyzed again and the most potent cytotoxic fraction and its IC50 were determined. Results: Statistical analysis showed that butanol extract of A. austroiranicum showed the most potent cytotoxic effects against OVCAR-3, HeLa and HUVEC cell lines with IC50 values of 38 +/- 2, 56 +/- 1.4, and 60 +/- 3.5 mu g/mL, respectively. On the other hand, for 7 fractions resulting from fractionation of the butanol extract, MTT assay results showed that 6th fraction (F) was the most cytotoxic fraction with IC50 of 2.7 +/- 0.26 and 7.5 +/- 0.5 mu g/mL for OVCAR-3 and HeLa cancer cell lines, respectively. Primary evaluation of the fraction by TLC and NMR analysis suggested the steroidal saponins as the main constituents. Conclusion: Allium austroiranicum showed significant cytotoxic effects against ovarian cancer cell line especially fractions assumed to contain steroidal saponins. The fraction constituents have the potential of being strong cytotoxic agents and the isolation and identification of compounds are suggested.