黄芩苷通过p53介导的SLC7A11下调诱导胃癌细胞铁死亡  

Baicalin Induces Ferroptosis in Gastric Cancer Cells via p53-mediated SLC7A11 Down-regulation

作　　者：苑小龙 魏征[2] 张俊萍[2] 郑芮 晁瑞 蔡小平[2] YUAN Xiaolong;WEI Zheng;ZHANG Junping;ZHENG Rui;CHAO Rui;CAI Xiaoping(The Second Clinical Medical College,Henan University of Chinese Medicine,Zhengzhou 450000,China;Henan Academy of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第二临床医学院,郑州450000 [2]河南省中医药研究院,郑州450000

出　　处：《中国实验方剂学杂志》2023年第6期71-78,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81873290);河南省中医药科学研究专项(2019ZBJ28);河南省中医管理局拔尖人才项目(豫卫中医2021-15)。

摘　　要：目的:以人胃癌SGC-7901细胞为研究对象,通过在培养液中添加不同浓度黄芩苷(0、100、200、400μmol·L-1),探讨黄芩苷对胃癌细胞增殖的抑制作用及可能的作用机制。方法:黄芩苷处理SGC-7901细胞后,采用噻唑蓝(MTT)比色法检测黄芩苷对胃癌细胞的抑制作用;同时通过添加3-氨基-4-环己基氨基苯甲酸乙酯(Fer-1)来观察细胞在黄芩苷处理后细胞的存活;使用实时荧光定量聚合酶链反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测铁死亡相关基因的表达;MTT比色法和酶联免疫吸附法分别检测丙二醛(MDA)含量和谷胱甘肽(GSH)水平;分别运用过表达和小干扰核糖核酸(siRNA)的方式来观察肿瘤蛋白53(p53)/溶质载体家族7成员11(SLC7A11)通路在铁死亡调控中的作用。结果:与空白组比较,黄芩苷处理后SGC-7901细胞活性明显降低(P<0.05,P<0.01),具有浓度和时间依赖性。与黄芩苷组比较,Fer-1干预显著缓解了黄芩苷引起的SGC-7901细胞活性降低(P<0.01)。与黄芩苷组比较,Fer-1+黄芩苷组细胞中MDA含量及前列腺素内过氧化物合成酶2(PTGS2)mRNA和蛋白水平显著降低(P<0.01),GSH活性、谷胱甘肽过氧化酶4(GPX4)mRNA和蛋白水平均显著升高(P<0.01)。与空白组比较,黄芩苷组细胞中SLC7A11蛋白表达水平明显降低(P<0.05,P<0.01),具有浓度依赖性。与黄芩苷组比较,SLC7A11过表达细胞在黄芩苷处理后细胞中活性氧(ROS)水平和MDA含量显著降低(P<0.01),GSH活性显著升高(P<0.01)。与空白组比较,黄芩苷组细胞中p53荧光强度显著升高(P<0.01)。与黄芩苷组比较,转染p53 siRNA的细胞在黄芩苷处理后细胞中p53蛋白表达水平显著降低(P<0.01),SLC7A11表达水平显著升高(P<0.01)。结论:黄芩苷能够有效抑制胃癌细胞SGC-7901的增殖,其机制可能与其调控p53/SLC7A11介导的细胞铁死亡有关。Objective:To explore the inhibitory effect of different concentration of baicalin(0,100,200,400μmol·L-1)on the proliferation of human gastric cancer SGC-7901 cells and the underlying mechanism.Method:SGC-7901 cells were treated with baicalin.Then methyl thiazolyl tetrazolium(MTT)assay was employed to examine the inhibitory effect of baicalin on the cells.At the same time,ferrostatin-1(Fer-1)was added to observe the viability of cells after baicalin treatment.The expression of ferroptosis-related genes was detected by Real-time polymerase chain reaction(Real-time PCR)and Western blot.The content of malondialdehyde(MDA)and the level of glutathione(GSH)were detected respectively by MTT assay and enzyme-linked immunosorbent assay.The role of tumor protein 53(p53)/solute carrier family 7 member 11(SLC7A11)pathway in the regulation of ferroptosis was investigated respectively via overexpression and small interfering RNA(siRNA)methods.Result:Compared with the blank group,baicalin decreased the viability of SGC-7901(P<0.05,P<0.01)in a dose-and time-dependent manner.The intervention of Fer-1 significantly alleviated the decrease of SGC-7901 cell viability caused by baicalin(P<0.01).In addition,compared with the baicalin group,Fer-1+baicalin group showed decrease in MDA content and the mRNA and protein levels of prostaglandin-endoperoxide synthase 2(PTGS2)in the cells(P<0.01),and increase in GSH activity and mRNA and protein levels of glutathione peroxidase 4(GPX4)(P<0.01).The protein level of SLC7A11 in the baicalin group was decreased compared with that in the blank group(P<0.05,P<0.01)in a dose-dependent manner.Compared with the baicalin group,the reactive oxygen species(ROS)level and MDA content in SLC7A11-overexpressing cells were significantly decreased after baicalin treatment(P<0.01),and the GSH activity was significantly increased(P<0.01).The fluorescence intensity of p53 in the cells of the baicalin group was increased compared with that of the blank group(P<0.01).Compared with the baicalin group,the expression

关 键 词：黄芩苷 胃癌细胞 铁死亡 肿瘤蛋白53 溶质载体家族7成员11 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学] R2-031R285.5