丙酸钠通过调节GPX4抑制人胃癌细胞增殖并诱导细胞铁死亡  

Sodium propionate inhibits the proliferation of human gastric cancer cells and induces ferroptosis by regulating GPX4

作　　者：马维娜 马秀杰 张欣 刘英坤 宋薇 何迪 曲相珍 MA Weina;MA Xiujie;ZHANG Xin;LIU Yingkun;SONG Wei;HE Di;QU Xiangzhen(Department of Clinical Pharmacy Office of the Pharmacy,Chifeng College Affiliated Hospital,Chifeng 024005,In-ner Mongolia,China)

机构地区：[1]赤峰学院附属医院药学部,赤峰024005

出　　处：《医学研究与战创伤救治》2024年第12期1299-1307,共9页Journal of Medical Research & Combat Trauma Care

基　　金：内蒙古自治区卫生健康科技计划项目(202201548)。

摘　　要：目的探讨丙酸钠(SP)通过调节谷胱甘肽(GSH)过氧化酶4(GPX4)抑制人胃癌细胞增殖并诱导细胞铁死亡。方法将AGS、NCI-N87细胞和转染Vector、GPX4过表达质粒的NCI-N87细胞分为DMSO组、DMSO+SP组、铁死亡诱导剂(FIN56)组和FIN56+SP组。通过CCK-8试验考察细胞的活力变化,并测定乳酸脱氢酶(LDH)积累情况。采用PCR分析GPX4 mRNA表达和蛋白质印迹分析GPX4蛋白表达。25只雌性BALB/c裸鼠随机分成5组(每组n=5):对照组、SP组、FIN56治疗组、FIN56+SP治疗组和FIN56+SP+甲磺酸去铁胺(DFOM)组。各组小鼠于右上肢皮下注射150μL含3×10^(6)NCI-N87细胞的PBS,每隔3天测量肿瘤体积。在注射后的第21天处死小鼠,收集肿瘤组织,免疫组化分析4-羟基壬烯醛(4-HNE)、Ki-67表达。结果与DMSO组相比,FIN56组AGS细胞和NCI-N87细胞的细胞活力、GSH水平显著降低(P<0.01),LDH积累、ROS水平显著增加(P<0.01)。与FIN56组相比,FIN56+SP组AGS细胞和NCI-N87细胞的细胞活力、GSH水平进一步降低(P<0.05),LDH积累、ROS水平进一步增加(P<0.01)。RT-PCR结果证实,SP以剂量依赖的方式抑制NCI-N87细胞和NCI-N87细胞中GPX4的mRNA表达(P<0.05)。与Vector+FIN56+SP组相比,GPX4-oe+FIN56+SP组的细胞活力显著升高(P<0.01),LDH积累显著降低(P<0.01)。与对照组相比,SP组、FIN56治疗组、FIN56+SP治疗组的肿瘤体积、肿瘤重量、Ki-67评分均显著降低(P<0.05),4-HNE评分显著增加(P<0.05),并且FIN56+SP治疗组的肿瘤体积、肿瘤重量、Ki-67评分、4-HNE评分的变化程度较SP组、FIN56组进一步增加(P<0.05)。与FIN56+SP治疗组相比,FIN56+SP+DFOM组的肿瘤体积、肿瘤重量、Ki-67评分显著增加(P<0.05),4-HNE评分显著降低(P<0.05)。结论SP可以通过下调GPX4的表达来促进胃癌细胞对FIN56诱导的铁死亡的敏感性。Objective The study aims to investigate the effect of sodium propionate(SP)on inhibiting the proliferation of human gastric cancer cells and inducing ferroptosis by regulating glutathione peroxidase 4(GPX4).Methods AGS,NCI-N87 cells and NCI-N87 cells transfected with Vector and GPX4 overexpression plasmid were divided into DMSO group,DMSO+SP group,ferroptosis inducer(FIN56)group and FIN56+SP group.CCK-8 test was used to investigate the changes of cell viability and determine the accumulation of lactate dehydrogenase(LDH).The expression of GPX4 mRNA was analyzed by PCR and GPX4 protein was analyzed by Western blot.25 female BALB/c nude mice were randomly divided into five groups(n=5 in each group):control group,SP group,FIN56 treatment group,FIN56+SP treatment group and FIN56+SP+deferoxamine mesylate(DFOM)group.Mice in each group were subcutaneously injected with 150μL PBS containing 3×10^(6)NCI-N87 cells in the right upper limb,and the tumor volume was measured every 3 days.The mice were killed on the 21st day after injection,and the tumor tissues were collected,and the expressions of 4-hydroxynonenal(4-HNE)and Ki-67 were analyzed by immunohistochemistry.Results Compared with DMSO group,the cell viability and GSH level of AGS cells and NCI-N87 cells in FIN56 group decreased significantly(P<0.01),while LDH accumulation and ROS level increased significantly(P<0.01).Compared with FIN56 group,the cell viability and GSH level of AGS cells and NCI-N87 cells in FIN56+SP group further decreased(P<0.05),while LDH accumulation and ROS level further increased(P<0.01).RT-PCR results confirmed that SP inhibited the expression of GPX4 mRNA in NCI-N87 cells and NCI-N87 cells in a dose-dependent manner(P<0.05).Compared with Vector+FIN56+SP group,the cell viability of GPX4-oe+FIN56+SP group increased significantly(P<0.01),and LDH accumulation decreased significantly(P<0.01).Compared with the control group,the tumor volume,tumor weight and Ki-67 score in SP group,FIN56 treatment group and FIN56+SP treatment group decreased signi

关 键 词：丙酸钠 短链脂肪酸 胃癌 铁死亡 FIN56 

分 类 号：R735.2[医药卫生—肿瘤]