Silver Nanoparticles Synthesized Coating with <i>Zataria Multiflora</i> Leaves Extract Induced Apoptosis in HeLa Cells Through p53 Activation.

Abstract

The biosynthesis of nanoparticles is widely considered today. This investigation was aimed at the biosynthesis and coating of Ag.NPs with <i>Zataria multiflora</i> (<i>Zm-</i>Ag.NPs) leaf extract and assessment of its apoptosis promoting effects. The <i>Zm</i>-Ag.NPs was characterized by UV-visible and FTIR spectroscopy, TEM, EDS, DLS, and measurement of zeta-potential. Apoptosis induction effects of <i>Zm-</i>Ag.NPs were assessed using acridine orange - propidium iodide <b>(</b>AO/PI), DAPI staining, caspase3/9 activation assay, and annexinV/PI assay. Changes in P53, matrix metalloproteinases 2 (MMPs), and vascular endothelial growth factor A (VEGF-A) genes expression were also assessed with semi-quantitative RT-PCR. The UV-visible spectroscopy results showed that the surface plasmon resonance band (SRP) for <i>Zm</i>-Ag.NPs was about 440 nm, also, FTIR spectroscopy indicated that plant material embedded around <i>Zm</i>-Ag.NPs. The TEM images of the samples revealed that the Ag.NPs varied in morphology and also, the presence of silver element was monitored with EDS. The mean size of <i>Zm</i>-Ag.NPs was 30 nm. The <i>Zm</i>-Ag.NPs reduced cell viability in a dose and time dependent manner (IC₅₀ = 15 μg/mL). AO/PI and DAPI staining indicated chromatin fragmentation and annexinV externalization assay using flow cytometer, confirmed promotion of programmed cell death in the treated cells. Apoptosis was induced through caspase 3/9 activation pathway. This promotion of apoptosis effects is not related with P53 gene up regulation. Finally, it was found that <i>Zm</i>-Ag.NPs inhibited cancer cell metastasis through a decrease in MMP and VEGFA expression. <i>Zm</i>-Ag.NPs acts as carrier of the plant material compound, and can be applied as anticancer agents.