Chemosensitivity of MCF-7 cells to eugenol: release of cytochrome-c and lactate dehydrogenase

Rana Al Wafai,Julnar Usta,Omar El-Rifai,Warde El-Rabih,Meghri Katerji,Remi Safi,Marwan El Sabban

doi:10.1038/srep43730

Rana Al Wafai, Julnar Usta + Show 5 more

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https://doi.org/10.1038/srep43730

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Journal: Scientific Reports	Publication Date: Mar 1, 2017
Citations: 31	License type: open-access

Affiliation: American University of Beirut

Abstract

Phytochemicals have been extensively researched for their potential anticancer effects. In previous study, direct exposure of rat liver mitochondria to eugenol main ingredient of clove, uncoupled mitochondria and increased F0F1ATPase activity. In the present study, we further investigated the effects of eugenol on MCF-7 cells in culture. Eugenol demonstrated: a dose-dependent decrease in viability (MTT assay), and proliferation (real time cell analysis) of MCF-7 cells, (EC50: 0.9 mM); an increase in reactive oxygen species; a decrease in ATP level and mitochondrial membrane potential (MitoPT JC-1 assay); and a release of cytochrome-c and lactate dehydrogenase (Cytotoxicity Detection Kit PLUS) into culture media at eugenol concentration >EC50. Pretreatment with the antioxidants Trolox and N-acetyl cysteine partially restored cell viability and decreased ROS, with Trolox being more potent. Expression levels of both anti- and pro-apoptotic markers (Bcl-2 and Bax, respectively) decreased with increasing eugenol concentration, with no variation in their relative ratios. Eugenol-treated MCF-7 cells overexpressing Bcl-2 exhibited results similar to those of MCF-7. Our findings indicate that eugenol toxicity is non-apoptotic Bcl-2 independent, affecting mitochondrial function and plasma membrane integrity with no effect on migration or invasion. We report here the chemo-sensitivity of MCF-7 cells to eugenol, a phytochemical with anticancer potential.

Highlights

Phytochemicals have been extensively researched for their potential anticancer effects
Other studies have demonstrated a protective effect of eugenol against N-methyl-N-nitro-N-nitrosoguanidine induced gastric carcinogenesis in a rat model[20,21]; this protective effect was mediated by the inhibition of NF-κB and modulation of Bcl-2 proteins[20,21]
We investigated the effect of eugenol, the main ingredient in clove, on MCF-7 cells and examined whether it causes the reduction in viability

Summary

Introduction

Phytochemicals have been extensively researched for their potential anticancer effects. Pretreatment with the antioxidants Trolox and N-acetyl cysteine partially restored cell viability and decreased ROS, with Trolox being more potent Expression levels of both anti- and pro-apoptotic markers (Bcl-2 and Bax, respectively) decreased with increasing eugenol concentration, with no variation in their relative ratios. Studies of the effects of eugenol on various cancer cell lines have reported induction of apoptosis in human melanoma G361 cells mediated by activation of caspases 3 and 622; induction of apoptosis in mast cells through translocation of p53 to the mitochondria[23]; and a decrease in the mitochondrial membrane potential of human leukemia cells (HL60) by activating caspase 3, an increase in reactive oxygen species (ROS) and a reduction in thiol levels[24]. Eugenol modulated growth and cyclooxygenase-2 expression in human colon cancer HT-29 cells[25], exerted an antiproliferative effect on colon cancer cells (HCT-15 and HT-29)[26] and inhibited www.nature.com/scientificreports/

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