Experimental Study Of Cancer Cell Membrane-Camouflaged Curcumin Nanoparticles Targeted Trigger Of Ferroptosis In Gastric Cancer Cells

Objective: In this study,a curcumin drug delivery system(MSN-CUR@CM)was designed to enhance the water solubility,tumor targeting and anti-gastric cancer effect of curcumin.By investigating the anti-tumor effect of MSN-CUR@CM on human gastric cancer(GC)cells SGC7901 and MGC803 in vitro,and further exploring the molecular mechanism of ferroptosis induced by MSN-CUR@CM,we aim to provide scientific theoretical basis for its clinical application.Methods:1.Preparation,characterization and properties of MSN-CUR@CMMesoporous silica nanoparticles(MSNs)were synthesized and the cancer cell membrane(CM)was prepared using the repeated freeze-thaw method.By loading CUR and cloaking homologous cancer CM onto MSNs,we fabricated the DDS MSN-CUR@CM.The loading capacity(LC)and encapsulation efficiency(EE)of CUR loaded MSNs were detected by ultraviolet/visible(UV/Vis)spectrophotometer.The morphological of the MSN-CUR@CM was characterized using transmission electron microscopy(TEM)and scanning electron microscopy(SEM).The elemental mapping of the nanomaterials was performed using field-emission TEM.10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)was conducted to detect the encapsulation of the cell membrane.The drug release of MSN-CUR@CM in different p H solutions was detected.The cytotoxicity on HUVEC and the blood compatibility were detected to evaluate the biological safety of MSN-CUR@CM.The dynamic light scattering(DLS)and zeta potential of the NPs were determined using Nano-ZS instrument.CUR release experiment,toxicological evaluation and water solubility were also tested.The in vivo and in vitro tumor targeting of the NPs was evaluated by cell uptake assay and in vivo distribution assay in tumor-bearing nude mice.2.Anti-tumor effect test of MSN-CUR@CM in SGC 7901 and MGC 803 cellsCell viability of SGC 7901 and MGC 803 cells following exposure to different concentration(0–40 μM)of CUR for different time(24 h and 48 h)was assessed using the CCK-8 kit.Cell viability of SGC 7901 and MGC 803 cells with different treatments(PBS,CUR,MSN-CUR,and MSN-CUR@CM,20 μM equivalent dose of CUR)was assessed using the CCK-8 kit and the calcein/PI staining assay.3.Detection of ferroptosis in SGC7901 and MGC803 cells induced by MSN-CUR@CMSGC 7901 and MGC 803 cells in different groups was pretreated with Fer-1(5 μM)for 2 h,and the cell viability was assessed using the CCK-8 kit.The cells were collected24 h after different treatments.The content of MDA,GSH and intracellular iron was detected by colorimetry,and the content of intracellular ROS was detected by flow cytometry.4.Exploration of the mechanism of ferroptosis triggered by MSN-CUR@CM in gastric cancer cellsHO-1 and GPX4 levels of SGC7901 cells with different treatments were detected using WB assay and IF assay.Result:1.We fabricated MSN-CUR@CM with a size of approximately 76.40±2.76 nm and the average surface charge of-26.50±0.92 m V.MSN-CUR@CM showed high water solubility,biocompatibility.The acidic solution(p H=5.0)was more conducive to CUR release than the neutral solution(p H=7.4).MSN-CUR@CM penetrated the cells by endocytosis and exhibited tumor-targeting both in vitro and in vivo.2.CUR suppressed SGC7901 and MGC803 cell viability in a concentration-and time-dependent manner after 24 h or 48 h,while MSN-CUR@CM inhibited viability of SGC7901 and MGC 803 cells much better than CUR.3.Pretreatment with Fer-1 effectively suppressed the cell death induced by exposure to CUR,MSN-CUR,and MSN-CUR@CM.The MDA,ROS,and iron levels were all higher whereas GSH was lower in CUR,MSN-CUR,and MSN-CUR@CM groups than they were in the control group in both SGC7901 and MGC803 cells,while the MSN-CUR@CM group presented the maximum degree of differentiation compared with the control group.4.CUR,MSN-CUR and MSN-CUR@CM all decreased the expression of GPX4 and increased the expression of HO-1 in SGC7901 cells,while MSN-CUR@CM presented the strongest effect.Conclusions:1.MSN-CUR@CM we synthesized exhibited high water solubility,biocompatibility,and tumor-targeting.2.CUR and MSN-CUR@CM triggered ferroptosis in SGC7901 and MGC803 cells.3.By downregulating the expression of GPX4 and upregulating that of HO-1,CUR and MSN-CUR@CM induced ferroptosis through the canonical and noncanonical ferroptosis pathways.4.In SGC7901 and MGC803 cells,MSN-CUR@CM enhanced the anti-gastric cancer effect of CUR.