重楼皂苷通过p53/SLC7A11信号轴促进三阴性乳腺癌细胞铁死亡的机制研究  

Studies on Paris saponin promoting iron death in triple negative breast cancer cells through p53/SLC7A11 signal axis

作　　者：李昕[1] 栗东海[1] 高小明[2] 山院飞[3] 薛江 Li Xin;Li Dong-Hai;Gao Xiao-Ming;Shan Yuan-Fei;Xue Jiang(Department of Thyroid and Breast Surgery,Affiliated Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia 010050,China;Department of Traditional Chinese Medicine,Affiliated Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia 010050,China;Department of General Surgery,Tianjin First Central Hospital,Tianjin 300110,China;Department of Thyroid and Breast Surgery,Bayannur City Hospital,Bayannur,Inner Mongolia 015000,China)

机构地区：[1]内蒙古医科大学附属医院甲状腺乳腺外科,内蒙古呼和浩特010050 [2]内蒙古医科大学附属医院中医科,内蒙古呼和浩特010050 [3]天津市第一中心医院普通外科,天津300110 [4]内蒙古自治区巴彦淖尔市医院甲状腺乳腺外科,内蒙古巴彦淖尔015000

出　　处：《解放军医学杂志》2023年第1期58-63,共6页Medical Journal of Chinese People's Liberation Army

基　　金：北京医学奖励基金会项目(YXJL-2020-1116-0412)。

摘　　要：目的探讨重楼皂苷对三阴性乳腺癌细胞铁死亡的影响及其机制。方法用0、10、20、30、40、50μmol/L重楼皂苷处理三阴性乳腺癌MDA-MB-231细胞,采用CCK-8法检测细胞活性。取对数生长期MDA-MB-231细胞,随机分为对照组、重楼皂苷组(30μmol/L重楼皂苷处理12 h)、抑制剂组(2μmol/L Ferrostatin-1处理12 h)与重楼皂苷+抑制剂组(2μmol/L Ferrostatin-1处理12 h后,30μmol/L重楼皂苷处理12 h),采用CCK-8法检测细胞活性,流式细胞仪检测细胞凋亡率,qRT-PCR和Western blotting检测二价金属离子转运体1(DMT1)、转铁蛋白受体1(TFR1)、谷胱甘肽过氧化物酶4(GPX4)、p53、p53/溶质运载蛋白7家族成员11(SLC7A11)的表达情况。结果CCK-8法检测结果显示,MDA-MB-231细胞存活率随重楼皂苷浓度的升高而降低,且呈剂量依赖性(P<0.05)。与对照组比较,重楼皂苷组细胞存活率以及CPX4、SLC7A11 mRNA和蛋白相对表达量降低,细胞凋亡率以及TFR1、DMT1、p53 mRNA和蛋白相对表达量升高,抑制剂组CPX4、SLC7A11 mRNA和蛋白相对表达量升高,细胞凋亡率以及TFR1、DMT1、p53 mRNA和蛋白相对表达量降低(P<0.05);与重楼皂苷组比较,重楼皂苷+抑制剂组细胞存活率以及CPX4、SLC7A11 mRNA和蛋白相对表达量升高,细胞凋亡率以及TFR1、DMT1、p53 mRNA和蛋白相对表达量降低(P<0.05);与抑制剂组比较,重楼皂苷+抑制剂组细胞存活率以及CPX4、SLC7A11 mRNA和蛋白相对表达量降低,细胞凋亡率以及TFR1、DMT1、p53 mRNA和蛋白相对表达量升高(P<0.05)。结论重楼皂苷可抑制三阴性乳腺癌细胞增殖,促进乳腺癌细胞铁死亡,其机制可能与p53/SLC7A11信号轴的调控作用有关。Objective To investigate the effect and mechanism of Paris saponins on iron death in triple negative breast cancer cells.Methods MDA-MB-231 cells were treated with 0,10,20,30,40 and 50μmol/L of Paris saponins.Cell viability was detected by CCK-8 method.MDA-MB-231 cells at logarithmic growth stage were randomly divided into control group,Paris saponin group(treated with 30μmol/L Paris saponin for 12 h),inhibitor group(treated with 2μmol/L Ferrostatin for 12 h)and Paris saponin+inhibitor group(treated with 30μmol/L Paris saponin for 12 h after treatment with 2μmol/L Ferrostatin-1 for 12 h),cell viability was detected by CCK-8 method,cell apoptosis rate was detected by flow cytometry.The expressions of divalent metal ion transporter 1(DMT1),transferrin receptor 1(TFR1),glutathione peroxidase 4(GPX4),p53 and p53/SLC7A11 were detected by qRT-PCR and Western blotting.Results CCK-8 assay showed that the survival rate of MDA-MB-231 cells was decreased with the increase of the concentration of Paris saponin in a dose-dependent manner(P<0.05).Compared with control group,the cell survival rate,the relative expression levels of CPX4,SLC7A11 mRNA and protein decreased,while of apoptosis rate,the relative,expression levels of TFR1,DMT1,p53 mRNA and protein increased in Paris saponin group,the relative expression levels of CPX4,SLC7A11mRNA and protein increased in inhibitor group,the apoptosis rate and the relative expressions of TFR1,DMT1 and p53 mRNA and protein were decreased(P<0.05).Compared with inhibitor group,the cell survival rate and the mRNA and protein relative expressions of CPX4 and SLC7A11 decreased,while the apoptosis rate and the mRNA and protein relative expressions of TFR1,DMT1 and p53 increased in parisaponin+inhibitor group(P<0.05).Conclusion Paris saponin inhibits the proliferation of triple negative breast cancer cells and promotes iron death in cancer cells,possibly through the regulation of p53/SLC7A11 signal axis.

关 键 词：三阴性乳腺癌 重楼皂苷 铁死亡 p53/SLC7A11信号轴 

分 类 号：R737.9[医药卫生—肿瘤]