Cytotoxic effect of NiFe2O4@Ag nanoparticle on adenocarcinoma gastric cell line (AGS) and assessment of the expression of CASP8, BAX, NRF2, and BCL2 genes

Abstract

In this research, the researchers evaluated the apoptotic property of NiFe2O4@Ag nanoparticle biosynthesized by Chlorella vulgaris on adenocarcinoma gastric cell line (AGS). The specification of synthesized NiFe2O4@Ag nanoparticles was investigated via several analytical analyses including, SEM (Scanning Electron Microscopy), TEM (Transmission Electron Microscopy), FTIR (Fourier Transform Infrared Spectroscopy), EDX (Energy Dispersive X-ray), Zeta potential analysis, DLS (Dynamic Light Scattering) and XRD (X-Ray Diffraction). To evaluate the in vitro cytotoxic effect, the AGS cells (cancer line) and HEK293 cells (normal cell line) were treated with different concentrations of NiFe2O4@Ag nanoparticles by MTT test. Moreover, the apoptogenic impact of the nanoparticles was assessed using Hoechst staining, annexin V- Fluorescein isothiocyanate (FITC) and propidium iodide (PI) staining, and Caspase 3 activity assay. Microscopic inspections of nanoparticles showed rounded shape and small particles in a size range of 30 to 60 nm. The MTT test indicated that the nanoparticles caused a dose-dependent inhibition of AGS cells (IC50 = 122.57 μg/mL). According to the Annexin V/PI staining assay, an increase in the frequency of apoptotic cells (51.75 %) in comparison to control cells was found. Based on the Caspase3 activity assay, nanoparticles led to an increase in Caspase3 level by 3.1 folds. Investigation of ROS (Reactive Oxygen Species) production exhibited that the nanoparticles can exert toxicity through an increase in ROS level. Cell cycle analysis confirmed an increase in the number of the cells in Sub-G1 step among nanoparticles treated cells. Furthermore, the expression of the BAX and CASP8 genes increased by 2.6 and 3.7 folds, respectively, whereas the BCL-2 gene was downregulated by 0.85 folds. Moreover, Hoechst staining indicated in vitro anti-tumor effect of the nanoparticles by appearance of apoptotic symptoms in AGS cells. Our results showed that NiFe2O4@Ag nanoparticled could efficiently cause the obstruction of AGS cell proliferation and apoptosis induction.