Spondias pinnata (L.f.) Kurz Leaf Extract Derived Zinc Oxide Nanoparticles Induce Dual Modes of Apoptotic-Necrotic Death in HCT 116 and K562 Cells

This study evaluates the effects of phyto-derived zinc oxide nanoparticles (ZnONPs) on human cancer cells, colon carcinoma HCT 116, and chronic myelogenous leukemic K562, along with normal lymphocytes/erythrocytes. The commercial, chemically synthesized ZnONPs (cZnONPs) were also assessed in parallel. Using an eco-friendly approach devoid of harmful chemicals, biogenic nanoparticles were synthesized from aqueous leaf extract of Spondias pinnata (SpLZnONPs) by a sol-gel method. Optical, structural, and elemental characterization of both particle types were carried out deploying UV-Vis/photoluminescence spectroscopy, FTIR, XRD, FESEM, HRTEM, and EDX. Both SpLZnONPs and cZnONPs displayed hexagonal wurtzite structure with particle sizes averaging 30 and 48.5 nm, respectively. SpLZnONPs were found to be cytotoxic to both cancer cell types while cZnONPs exhibited toxicity only against HCT 116 cells. Interestingly, the cytomorphological changes and analysis of DNA laddering pattern observed in these treated cells were indicative of simultaneous induction of dual modes of death involving apoptosis and necrosis. Flow cytometric analysis of cell-cycle distribution, clonogenic, wound healing, and comet assays provided evidences of the antiproliferative potential of the tested nanoparticles. Apoptosis induction via oxidative stress-mediated Ca²⁺ release, ROS generation, loss of mitochondrial membrane potential, and externalization of phosphatidylserine was also determined biochemically. Relative expression of apoptotic genes was quantified using RT-qPCR and Western blot analysis. Mitotic index analysis, MTT, and hemolytic assays on lymphocytes and erythrocytes clearly revealed the absence of any deleterious effect(s) of SpLZnONPs in these cells compared with the toxicity of the chemically derived cZnONPs, thereby attesting to the biocompatibility and selective action of the biogenic nanoparticles.

Keywords: Apoptosis; HCT 116; K562; Necrosis; Spondias pinnata; Zinc oxide nanoparticles.