Ethyl Acetate Fraction of Anethum graveolens Seeds Exerts an Antiproliferative Effect by Inhibiting Anti‑apoptotic Proteins in MCF‑7 and PC‑3 Cells: An in vitro and Molecular Docking Study

Abstract

Objectives: We investigated, the antiproliferative effect of ethyl acetate fraction of dill (EAFD), on MCF‑7 and PC‑3 cell lines and its two most active components, anethole and carvone by molecular docking analysis.Materials and Methods: In‑vitro assays, like cell viability assay and measurement of reactive oxygen species, were performed besides performing Giemsa stain and other fluorescent stains; JC‑1 dye, dual mixed stain ethidium bromide/acridine orange and 4,6‑diamidino‑2‑phenylindole stain to study morphological characteristics, including molecular docking analysis. EAFD concentrations (0.2, 0.4, 0.6, 0.8, 1.0 mg/ml) were used.Results: The EAFD prominently inhibited the proliferation of MCF‑7 cells and PC‑3 cells by dose‑dependent and time‑dependent methods. Increased exposure of EAFD to MCF‑7 and PC‑3 cells increases the level of intracellular oxidative stress. Similarly, EAFD exposure confirms the morphological alternations such as cell shrinkage, membrane disruption, nuclear condensation, and blebbing in phase‑contrast microscopy and even fluorescent microscopy stains can lead to mitochondrial membrane degradation, chromatin condensation, nuclear fragmentation. Analyses of docking results suggest that anethole and carvone bind to the hydrophobic patches of Bcl‑2 and Bcl-xL through hydrophobic interactions.Conclusion: The EAFD may be antiproliferative activity and leading to pro‑apoptotic cell death. Molecular docking analysis of Bcl‑2 and Bcl-xL anti‑apoptotic protein indicated that the antiproliferative activity and pro‑apoptotic cell deaths by EAFD are possibly due to inhibition of these proteins.

Keywords: Anethum graveolens; anti‑proliferation; apoptosis; cancer; herbal medicine; intracellular oxidative stress; molecular docking