陆军军医大学学报 (Jul 2024)
Sodium-hyaluronate-modified calcium peroxide nanoparticles induce pyroptosis in gastric cancer cells in vitro
Abstract
Objective To investigate the role of sodium-hyaluronate-modified calcium peroxide nanoparticles (SH-CaO2 NPs) in inducing pyroptosis in human gastric cancer cells and its possible mechanisms. Methods Transmission electron microscopy (TEM), X-ray diffraction (XRD), infrared spectroscopy, and zeta potential test were used to confirm the synthesis of SH-CaO2 NPs. Cell scratch assay and CCK-8 assay were employed to observe the impacts of SH-CaO2 NPs on the migration and proliferation of human gastric cancer cell line HGC-27. The generation of reactive oxygen species (ROS) was observed with confocal laser scanning microscopy (CLSM) and quantified with flow cytometry in the cells after SH-CaO2 NPs treatment or with pretreatment with ROS inhibitor NAC. Furthermore, the effects of pretreatment of NLRP3 inhibitor (MCC950) and Caspase-1 inhibitor (VX765) on the proliferative activity and on expression of their own and their downstream GSDMD in HGC-27 cells were also investigated with CCK-8 assay, immunofluorescence assay and Western blotting. Results TEM images, XRD, infrared spectroscopy, and zeta potential test confirmed the successful preparation of SH-CaO2 NPs. Cell scratch assay and CCK-8 assay showed that application of SH-CaO2 NPs for 24 h significantly inhibited the proliferation of HGC-27 cells (P < 0.001), while, CLSM and flow cytometry indicated the treatment also promoted the production of ROS (P < 0.001). Pretreatment of ROS inhibitor NAC resulted in up-regulation of NLRP3, and increased expression levels of cleaved Caspase-1 and N-terminal fragment of GSDMD (P < 0.001), while pretreatment of both NLRP3 inhibitor and Caspase-1 inhibitor could reverse the process. Conclusion SH-CaO2 NPs inhibit cell viability of human gastric cancer, which may mediate the inflammatory response and pyroptosis by activating the ROS/NLRP3/Caspase-1/GSDMD signaling pathway.
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