白藜芦醇通过调控二氢乳清酸脱氢酶介导的铁死亡增加胰腺癌细胞顺铂敏感性  

Resveratrol Increases Cisplatin Sensitivity in Pancreatic Cancer Cells by Modulating Dihydroorotatede Hydrogenase-Mediated Ferroptosis

作　　者：郑艳霞 吕思懿 李婷婷 丁玉松[1,2] ZHENG Yanxia;LÜSiyi;LI Tingting;DING Yusong(School of Medicine,Shihezi University,Shihezi 832061,China;School of Public Health,Xinjiang Medical University,Ürümqi 830000,China)

机构地区：[1]石河子大学医学院,新疆石河子832061 [2]新疆医科大学公共卫生学院,新疆乌鲁木齐830000

出　　处：《食品科学》2025年第9期189-205,共17页Food Science

基　　金：新疆医科大学医学科学研究所项目(YXYJ20240103)。

摘　　要：目的:探讨白藜芦醇(resveratrol,RSV)通过抑制二氢乳清酸脱氢酶(dihydroorotatede hydrogenase,DHODH)增加胰腺癌细胞对顺铂(cisplatin,DDP)敏感性的效应,并从铁死亡角度分析其潜在的作用机制。方法:使用不同剂量的DDP处理人胰腺癌细胞系PANC1(0、10、20、30、40、50、60、70、80μmol/L)和人原位胰腺腺癌细胞系BxPC-3(0、2、4、6、8、10、12、14μmol/L),诱导细胞铁死亡。使用布喹那(brequinar,BQR)和RSV对细胞进行干预,探讨其对DDP诱导的胰腺癌细胞铁死亡的影响。通过测定细胞活力、细胞中谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、溶质载体家族7成员11 xCT和DHODH蛋白的表达水平,探讨DDP及BQR/RSV联合干预对细胞内铁死亡相关分子的影响。通过分光光度法测定丙二醛(malondialdehyde,MDA)含量,评估脂质过氧化程度,并且通过检测细胞内活性氧(reactive oxygen species,ROS)的水平,进一步了解氧化应激的变化。通过JC-1荧光探针检测线粒体膜电位(mitochondrial membrane potential,MMP),探讨细胞内线粒体功能的变化。采用免疫荧光法检测细胞增殖标志物Ki-67的水平,评估细胞增殖的变化。结果:DDP处理诱导胰腺癌细胞发生铁死亡(P<0.05)。BQR和RSV单独使用或联合DDP作用均能够显著降低胰腺癌细胞中GPX4、xCT和DHODH的蛋白表达水平(P<0.05),同时降低MMP和Ki-67水平(P<0.05)。相反,BQR/RSV处理提高了细胞内ROS和MDA水平,与DDP联用的效果更为显著(P<0.05)。转染DHODH腺病毒后,显著逆转了DDP+RSV联合处理对胰腺癌细胞增殖抑制的作用。结论:在胰腺癌组织中DHODH表达上调。DHODH抑制能够促进胰腺癌细胞铁死亡,增强顺铂敏感性。RSV可能作为一种DHODH的抑制剂促进DDP诱导的胰腺癌细胞铁死亡。Objective:To investigate the effect of resveratrol(RSV)in increasing the sensitivity of pancreatic cancer cells to cisplatin(DDP)through inhibition of dihydroorotatede hydrogenase(DHODH)and to analyze its potential mechanism of action from the perspective of ferroptosis.Methods:The human pancreatic ductal cancer cell line PANC1(0,10,20,30,40,50,60,70,and 80μmol/L)and the human orthotopic pancreatic cancer cell line BxPC-3(0,2,4,6,8,10,12,and 14μmol/L)were treated with different doses of DDP to induce ferroptosis.Subsequently,the effect of intervention with brequinar(BQR)or RSV was explored on DDP-induced ferroptosis in pancreatic cancer cells.To explore the effects of DDP combined with BQR or RSV on ferroptosis-related intracellular molecules,cell viability was determined by the cell counting kit-8(CCK-8)assay,and Western blot was used to detect the expression levels of glutathione peroxidase 4(GPX4),recombinant solute carrier family 7 member 11(SLC7A11 or xCT)and DHODH in cells.The content of malondialdehyde(MDA)was determined spectrophotometrically to evaluate the degree of lipid peroxidation.The level of intracellular reactive oxygen species(ROS)was detected using DCFH-DA probe to further understand the changes of oxidative stress.The fluorescent probe JC-1 was used to detect mitochondrial membrane potential(MMP)to explore the changes of mitochondrial function in cells.Immunofluorescence was used to detect the level of the cell proliferation marker Ki-67 to evaluate the changes of cell proliferation.Results:DDP treatment induced ferroptosis in pancreatic cancer cells(P<0.05).Both BQR and RSV alone or in combination with DDP significantly decreased the protein expression levels of GPX4,xCT,and DHODH in pancreatic cancer cells(P<0.05),as well as the levels of MMP and Ki-67(P<0.05).In contrast,BQR/RSV treatment elevated the levels of intracellular ROS and MDA(P<0.05),and the effect was more pronounced in combination with DDP(P<0.05).Transfection with DHODH adenovirus significantly reversed the inhibitory effect

关 键 词：胰腺癌 铁死亡 顺铂 二氢乳清酸脱氢酶 白藜芦醇 

分 类 号：R151.3[医药卫生—营养与食品卫生学]