Abstract
BackgroundEugenol is a natural phenolic compound and possesses anticancer and antibacterial activities. Breast cancer is a major global health problem, and most of the chemotherapeutic agents are highly toxic with long-term side effects. Therefore, this study aimed to explore the possibility of using eugenol as an anti-metastatic and anti-proliferative agent against MDA-MB-231 and SK-BR-3 breast cancer cells.MethodsBreast cancer cell lines MDA-MB-231 and SK-BR-3 were treated with eugenol and cell proliferation was measured using a real-time cell electronic sensing system. Annexin V analysis with flow cytometry was used to detect the effect of eugenol on cell death. In MDA-MB-231 and SK-BR-3 cells, metastatic potential after eugenol treatment was examined using a wound-healing assay. Real-time PCR was used to study the effect of eugenol on the expression of anti-metastatic genes such as MMP2, MMP9, and TIMP-1, and genes involved in apoptosis including Caspase3, Caspase7, and Caspase9.ResultsTreatment with 4 μM and 8 μM eugenol for 48 h significantly inhibited cell proliferation of MDA-MB-231, with an inhibition rate of 76.4%, whereas 5 μM and 10 μM of eugenol for 48 h significantly inhibited the proliferation of SK-BR-3 cells with an inhibition rate of 68.1%. Eugenol-treated cells showed significantly decreased MMP2 and MMP9 expression and an insignificant increase in TIMP1 expression in HER2 positive and triple negative breast cancer cells. Eugenol significantly increased the proportion of MDA-MB-231 and SK-BR-3 cells in late apoptosis and increased the expression of Caspase3, Caspase7, and Caspase9.ConclusionTo the best of our knowledge, this is the first study to describe the anti-metastatic effect of eugenol against MDA-MB-231 and SK-BR-3 breast cancer cell lines.
Highlights
Eugenol is a natural phenolic compound and possesses anticancer and antibacterial activities
Effect of eugenol on cell viability To determine the anti-proliferative effect of different eugenol concentrations against MDA-MB-231 and SK-BR-3 cell growth, the cell proliferation assay was performed using the MTT method and real-time cell analysis
Incubation with 12 μM eugenol for 24 and 48 h caused a significant decrease in SK-BR-3 cell proliferation by 66.7 and 67.3%, respectively, compared to untreated cells (Fig. 2b)
Summary
Eugenol is a natural phenolic compound and possesses anticancer and antibacterial activities. Breast cancer is a major global health problem, and most of the chemotherapeutic agents are highly toxic with long-term side effects. Breast cancer is a major health problem with high morbidity and mortality rates among women worldwide [1]. Its high prevalence and differences in response patterns to various treatment modalities and clinical outcomes provides a strong rationale for identifying natural and synthetic cancer preventive agents [2]. Metastasis is considered one of the main causes of death [8]. There was a link between the high levels of a group of matrix metalloproteinases (MMPs), a family of 23 structurally and functionally related endopeptidases [10], and most human tumor cell lines [11]. The level MMP-2 and MMP-9 are overexpressed [13] which is associated with a shortened relapse-free survival [16]
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