Silver nanoparticles Synthesized coating with Leaves<i> Zataria multiflora</i> extract Induced Apoptosis in HeLa cells through p53 activation

Abstract

Aims: The biosynthesis of nanoparticles is widely considered today. The aim of this investigation was the biosynthesis and coating of Ag.NPs with Zataria multiflora (Zm-Ag.NPs) leaf extract and assess its apoptosis promoting effects. Methods: 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) and DAPI staining, caspase3/9 activation assay and annexinV/PI assay. We also assess changing on P53, matrix metalloproteinases 2 (MMPs) and vascular endothelial growth factor A (VEGF-A) genes expression with semi-quantitative RT-PCR. Results: The UV-visible spectroscopy results showed that the surface plasmon resonance band (SRP) for Zm-Ag.NPs was around 440 nm also FTIR spectroscopy indicated plant material embedded around of Zm-Ag.NPs. The TEM images of the samples revealed that the Ag.NPs varied in morphology also presence of the silver element was monitories with EDS. the mean size of Zm-Ag.NPs were 30 nm. The Zm-Ag.NPs decrease cell viability in a dose and time dependent manner (IC50 = 15 μg/ml). AO/PI and DAPI staining indicated chromatin fragmentation also annexinV externalization assay using flow cytometer confirm promoting programmed cell death in treated cells. Apoptosis induced through caspase 3/ 9 activation pathway. This promoting apoptosis effects didn't relate 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. Conclusion: Zm-Ag.NPs acts as carrier for plant material compound that can apply as anticancer agents.