Evaluation of the effect of iron oxide nanoparticles functionalized by glucose and conjugated with coumarin (Fe3O4@Glu-Coumarin NPs) on the expression of CASP8, CASP9, p53, mTOR1, and MAPK1 genes in liver cancer cell line

Abstract

Due to the high morbidity and mortality of liver cancer, many researchers are seeking novel anticancer formulations. Complex iron oxide containing compounds could be used for site directed drug delivery using a magnetic force. Due to the anticancer property of Coumarin, this study aimed to study the effect of iron oxide nanoparticles functionalized by glucose and conjugated with Coumarin (Fe3O4@Glu-Coumarin NPs) on the expression of cell cycle and apoptosis regulating genes in a liver cancer cell line. Fe3O4@Glu-Coumarin NPs were synthesized and their physicochemical properties were evaluated by FT-IR, XRD (X-Ray diffraction), EDS mapping, and electron microscope imaging. Anti-proliferative activity of the NPs on HepG2 cells was studied by MTT assay. The effect of Fe3O4@Glu-Coumarin NPs on the expression of CASP8, CASP9, p53, mTOR1, and MAPK1 genes was investigated by quantitative PCR. Moreover, the apoptogenic impact of the Fe3O4@Glu-Coumarin NPs was assessed using annexin V- Fluorescein isothiocyanate (FITC) - propidium iodide (PI) staining and cell cycle analysis. The synthesized NPs were spherical, with a moderate level of aggregation and in a size range of 26–46 nm. FT-IR, XRD, and EDS mapping confirmed the proper synthesis and purity of the synthesized NPs. The zeta potential value of the NP was −23 mV. According to the MTT assay, Fe3O4@Glu-Coumarin NPs showed a considerable anti-proliferative effect on liver cancer cells, and the 50 % inhibitory concentration was determined 120 μg/mL. Quantitative PCR assay showed that the NPs significantly increased the expression of CASP8, p53, and MAPK1 genes by 1.94, 4.87, and 3.87 folds, respectively, while the mTOR1 gene was reduced by −6.33 folds. The expression of the CASP9 had an insignificant reduction by 0.06 folds. Annexin V/PI staining assay showed an increase in frequency of apoptotic cells in comparison to control cells. Cell cycle analysis confirmed an increase in the number of the cells in G0/G1step among nanoparticles treated cells. Our results showed that Fe3O4@Glu-Coumarin NPs could induce cell apoptosis by interfering with the expression of the cell regulatory genes.