Regulation of solute carrier SLC3A2 in ferroptosis of SH-SY5Y cells
Objective To investigate the regulation of SLC3A2 on ferroptosis in human neuroblastoma SH-SY5Y cells.Methods The SH-SY5Y cell line was used to construct cell models of SLC3A2 gene knockdown(SLC3A2KD),SLC3A2 gene overexpression(SLC3A2OE),and knockdown with subsequent overexpression(SLC3A2KD+OE)Normal SH-SY5Y cells,SLC3A2KD,SLC3A2OE,and SLC3A2KD+OE cells were treated separately with Erastin,a ferroptosis-specific inducer.Cell viability was measured by the CCK8 method.Malondialdehyde(MDA)content,glutathione(GSH)content,glutathione peroxidase(GSH-Px)activity,and tissue iron content was determined using assay kits.Lipid ROS levels were detected by flow cytometry,while SLC7A11 and ACSL4 protein expressions were measured by Western blot.Results After treated with Erastin,compared to the Erastin group,the SLC3A2KD+Erastin group had decreased cell viability,GSH content,GSH-Px activity,and SLC7A11 protein expression level(F=44.576,F=32.487,F=56.562,F=118.709,P＜0.01),and increased lipid ROS,MDA,cellular iron,and ACSL4 protein expression level(F=77.392,F=53.256,F=79.710,F=75.027,P＜0.01).In contrast,the SLC3A2OE+Erastin and SLC3A2KD+OE+Erastin groups saw restored cell viability(F=47.885,P＜0.01;F=25.003,P＜0.05),increased GSH content,GSH-Px activity,and SLC7A11 protein expression level(F=35.438,F=37.974,F=26.520,P＜0.01;F=70.271,F=36.666,F=25.045,P＜0.01),and reduced lipid ROS,MDA,cellular iron,and ACSL4 protein expression level(F=178.026,F=83.256,F=93.156,F=34.907,P＜0.01;F=164.425,F=19.157,F=180.557,F=43.385,P＜0.01).Conclusion SLC3A2 might increase the sensitivity of SH-SY5Y cells to Erastin-induced ferroptosis by affecting the stability of the SLC7A11 protein.
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