溶质转运蛋白SLC3A2在SH-SY5Y细胞铁死亡中的调控作用  

Regulation of solute carrier SLC3A2 in ferroptosis of SH-SY5Y cells

作　　者：佟晓敏 李朝菲 吕懿[1] 郑金平[1,2] TONG Xiao-min;LI Zhao-fei;LV Yi;ZHENG Jin-ping(Department of Toxicology,School of Public Health,Shanxi Medical University,Taiyuan,Shanxi 030001,China;Department of Public Health and Preventive Medicine,Changzhi Medical College,Changzhi,Shanxi 046000,China)

机构地区：[1]山西医科大学公共卫生学院卫生毒理学教研室,山西太原030001 [2]长治医学院公共卫生与预防医学系,山西长治046000

出　　处：《毒理学杂志》2024年第2期101-108,共8页Journal of Toxicology

基　　金：山西省重点研发计划(国际合作,201703D421021);山西省“1331工程”提质增效项目(2021-5-2-2-B1);山西省基础研究计划(自由探索类)青年科学研究项目(20210302124301);长治医学院科技创新团队(CX202001)。

摘　　要：目的探讨SLC3A2对人神经母细胞瘤SH-SY5Y细胞铁死亡的调控作用。方法使用SH-SY5Y细胞株构建SLC3A2基因敲低细胞模型SLC3A2^(KD),SLC3A2基因过表达细胞模型SLC3A2^(OE),及敲低后过表达细胞模型SLC3A2^(KD+OE)。用铁死亡特异性诱导剂Erastin分别处理正常SH-SY5Y细胞、SLC3A2^(KD)细胞、SLC3A2^(OE)细胞和SLC3A2^(KD+OE)细胞。应用CCK8法检测细胞存活率,试剂盒法检测丙二醛(MDA)含量、谷胱甘肽(GSH)含量、谷胱甘肽过氧化物酶(GSH-Px)活力和细胞铁含量,流式细胞术检测脂质ROS水平,Western blot检测SLC7A11和ACSL4蛋白表达。结果使用Erastin处理各组细胞后,与Erastin组相比,SLC3A2^(KD)+Erastin组细胞存活率、GSH含量、GSH-Px活力和SLC7A11蛋白表达量降低(F=44.576,F=32.487,F=56.562,F=118.709,P<0.01),MDA含量、脂质ROS水平、细胞铁含量和ACSL4蛋白表达量增高(F=77.392,F=53.256,F=79.710,F=75.027,P<0.01);SLC3A2^(OE)+Erastin组和SLC3A2^(KD+OE)+Erastin组细胞存活率恢复(F=47.885,P<0.01;F=25.003,P<0.05),GSH含量、GSH-Px活力和SLC7A11蛋白表达量增高(F=35.438,F=37.974,F=26.520,P<0.01;F=70.271,F=36.666,F=25.045,P<0.01),MDA含量、脂质ROS水平、细胞铁含量和ACSL4蛋白表达量下降(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)。结论SLC3A2可能通过影响SLC7A11蛋白稳定性增加SHSY5Y细胞对Erastin诱发的铁死亡的敏感性。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(SLC3A2^(KD)),SLC3A2 gene overexpression(SLC3A2^(OE)),and knockdown with subsequent overexpression(SLC3A2^(KD+OE)).Normal SH-SY5Y cells,SLC3A2^(KD),SLC3A2^(OE),and SLC3A2^(KD+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 SLC3A2^(KD)+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 SLC3A2^(OE)+Erastin and SLC3A2^(KD+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.

关 键 词：SLC3A2 铁死亡 人神经母细胞瘤细胞 Erastin SLC7A11 

分 类 号：R114[医药卫生—卫生毒理学]