Silver Nanoparticles Synthesized Coating with Zataria Multiflora Leaves Extract Induced Apoptosis in HeLa Cells Through p53 Activation

Iran J Pharm Res. Spring 2018;17(2):627-639.

Abstract

The biosynthesis of nanoparticles is widely considered today. This investigation was aimed at the biosynthesis and coating of Ag.NPs with Zataria multiflora (Zm-Ag.NPs) leaf extract and assessment of its apoptosis promoting effects. The Zm-Ag.NPs was characterized by UV-visible and FTIR spectroscopy, TEM, EDS, DLS, and measurement of zeta-potential. Apoptosis induction effects of Zm-Ag.NPs were assessed using acridine orange - propidium iodide (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 Zm-Ag.NPs was about 440 nm, also, FTIR spectroscopy indicated that plant material embedded around Zm-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 Zm-Ag.NPs was 30 nm. The Zm-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 Zm-Ag.NPs inhibited cancer cell metastasis through a decrease in MMP and VEGFA expression. Zm-Ag.NPs acts as carrier of the plant material compound, and can be applied as anticancer agents.

Keywords: Apoptosis; MMP-9; Silver nanoparticle; VEGF-A; Zataria multiflora.