In vitro apoptosis and molecular response of engineered green iron oxide nanoparticles with l-arginine in MDA-MB-231 breast cancer cells

Abstract

Iron oxide nanoparticles (IONPs) have immense applications in pharmaceutical industries, diagnostics, therapeutics, and biomedicine. In this study, the aqueous leaf extract of Mimosa pudica was utilized for the synthesis of IONPs. The color change of the solution after adding 3 mM ferrous sulfate (FeSO4) confirmed biosynthesis of IONPs. The spherical IONPs were coated with l-arginine (cIONPs) and were found to be in the size range of 20 nm–48 nm, and toxicity analysis revealed IC50 value of these cIONPs were 67.69 μg/ml against MDA-MB-231 breast cancer cells and 156.28 μg/ml against normal 3T3 L1 cells. The apoptosis study revealed membrane blebbing, chromatin condensation and change in cell morphology and mitochondrial membrane potential assay confirmed mitochondrial membrane depolarization (Δψm) with 38.19% membrane damage. Dichlorodihydrofluorescein diacetate (H2DCFDA) analysis confirmed the production of reactive oxygen species (ROS) (57.95%) in the cIONPs treated breast cancer cells. Flow cytometry results confirmed that cell cycle got arrested at S (16.52%) stage, even better in G2/M stage (52.82%). Extensive DNA fragmentation in treated cells was detected via agarose gel electrophoresis. Quantitative reverse transcription PCR (RTqPCR) confirms the upregulation of endoplasmic reticulum (ER) stress XBPs, PERK, CHOP, ATF-6 genes, and downregulation of IRE1 alpha gene. The study showed cIONPs could be used as potential anticancer agent in biomedical and pharmaceutical applications.