Induction of Apoptosis by Ethanol, Methanol, and Ethyl Acetate Extracts from Cyperus Rotundus' Leaf on In Vitro Human Ovarian, Cervix, and Breast Cancer Cell Lines.

Abstract

Background: Cervical, breast, and ovarian cancers exhibit significant incidence and fatality rates, necessitating diverse approaches for effective cancer cell eradication while preserving normal cells. The aim of this study is to explore the in vitro apoptosis-inducing properties of hydroalcoholic extracts from Cyperus rotundus' (C. rotundus) leaf using human gynecological cancer cell lines.

Materials and methods: In this experimental study, Hydroalcoholic extracts were prepared from the leaf of C. rotundus using ethanol, methanol, and ethyl acetate. These extracts were applied to MCF-7, HeLa, OVCAR-3, and Vero cell lines at concentrations of 3.125, 6.25, 12.5, and 25 g/ml. MTT test, assessing inhibition of proliferation at 50% (IC₅₀), was employed to evaluate each extract's ability to inhibit cell proliferation. Subsequently, apoptosis-related gene and protein regulation were examined using reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.

Results: Gas chromatography-mass spectrometry (GC-MS) analysis revealed that the methanolic extract contained hexadecanoic acid and dodecanoic acid. The ethanolic extract was found to have norspermidine and desulphosinigrin. Additionally, the ethyl acetate extract included vitamin E, 1-heptatriacotanol, lupeol, betulin, stigmasterol, and stearic acid. The HeLa treatment group with 6.25 μg/ml of ethyl acetate extract, MCF-7, and OVCAR-3 cells with 3.125 μg/ ml of methanol extract treatment group exhibited the most significant growth inhibition in the MTT assay. Further analysis of these treatment groups revealed that the transcription and translation of BAX, Caspase-3, Caspase-8, and Caspase-9 increased overall, whereas Bcl-2 decreased in all cell lines.

Conclusion: Hydroalcoholic extracts from C. rotundus' leaf may enhance the apoptosis of cancer cells by modulating the transcription, translation, and post-translation of proteins, with minimal impact on the growth and survival of non-cancerous cells.