Eugenol–AgNPs Induce Apoptosis and Upregulate Apoptosis -Related Caspase Gene Expression in MCF-7/C3 Breast Cancer Cells

Abstract

The purpose of this study was to assess the anticancer potential of Eugenol–AgNPs obtained from the extract of Fagonia indica and find out its mode of action on MCF-7/C3 breast cancer cells. GC-MS analysis revealed a total of 48 components, with Eugenol accounting for 26.45% of the relative peak area. In all of the analyses performed using HPLC-DAD method, the Eugenol peak constituted 26.6% of the total summation of peak area. The purity attained in the Eugenol-enriched fraction is about 94.7 ± 1.5%. The characterization of Eugenol–AgNPs was carried out using various techniques such as FTIR, UV–Vis, TEM/HRTEM, DLS, XRD, EDX, and zeta potential. The TEM/HRTEM analysis confirmed the existence of nanoparticles mostly in spherical shape with average size being 44.1 ± 6.7 nm. The hydrodynamic diameter as observed by DLS was 68.6 ± 5.4 nm. Overall characterization confirmed formation of Eugenol–AgNPs and its physicochemical properties including zeta potential, which was measured as −24.8 ± 2.9 mV. The nanoparticles showed strong dose-dependent cytotoxic effects against MCF-7/C3 cells with IC₅₀ equal to 1.854 µg/mL. The analysis using Annexin V/PI demonstrated a significant increase in apoptotic cellsm while the RT-qPCR revealed increased in Caspase-3, Caspase-8, and Caspase-9 expression. Moreover, Eugenol-AgNPs has also been observed to have selectivity regarding the cytotoxicity of the MCF-7/C3 cells as compared to MCF-10A cells, where selectivity indexes of Eugenol-Ag noted to be 5.31. These collective data indicates that Eugenol-AgNPs have anti-breast cancer properties because of apoptosis induction. They appear to be a promising candidate for a plant-based nanocomposite that begs for more research to establish its clinical maximum