土荆皮乙酸上调HMOX1诱导宫颈癌Siha细胞凋亡和铁死亡  

The mechanistic analysis of Siha apoptosis and ferroptosis caused in cervical cancer cells by Pseudolaric acid B up-regulation of HMOX1

作　　者：董晶晶 石少卿 李玉琼 牛璐璐 孙瑞芬 DONG Jingjing;SHI Shaoqing;LI Yuqiong;NIU Lulu;SUN Ruifen(Yunnan University of Traditional Chinese Medicine,Yunnan Kunming 650500,China;The First Affiliated Hospital of Kunming Medical University,Yunnan Kunming 650032,China)

机构地区：[1]云南中医药大学,云南昆明650500 [2]昆明医科大学第一附属医院,云南昆明650032

出　　处：《现代肿瘤医学》2024年第18期3413-3419,共7页Journal of Modern Oncology

基　　金：云南省科技厅科技计划项目[编号:202201AT070043,2019FF002(-011)]。

摘　　要：目的:探讨土荆皮乙酸(Pseudolaric acid B,PAB)诱导宫颈癌Siha细胞发生凋亡和铁死亡的机制。方法:CCK8法检测PAB对宫颈癌Siha细胞的细胞毒性。Annexin V-FITC和PI双荧光染色检测细胞凋亡。PI和Hochest染色,荧光显微镜观察细胞核形态和细胞死亡情况。流式细胞仪检测细胞周期及细胞内活性氧(ROS)水平。脂质过氧化传感器BODIPY^(TM)581/591 C11染色,荧光显微镜检测细胞内脂质过氧化。JC-1染色,荧光显微镜观察细胞内线粒体膜电位变化。Western blot检测铁死亡相关蛋白GPX4、SLC7A11、Ferritin、HMOX1和凋亡相关蛋白PARP、c-IAP1、c-IAP2、XIAP、Survivin的表达。RT-PCR检测铁死亡相关基因的mRNA表达水平。结果:PAB能够显著抑制宫颈癌Siha细胞的增殖,促进细胞凋亡,阻滞细胞周期。其次,PAB促进细胞内ROS水平升高,脂质过氧化物积累,和线粒体膜电位上升。同时,上调HMOX1蛋白和mRNA的表达,下调GPX4、SLC7A11蛋白和mRNA表达,下调Ferritin、c-IAP1、c-IAP2、XIAP、Survivin的蛋白表达。结论:PAB可以显著促进宫颈癌Siha细胞的细胞死亡,作用机制可能与上调HMOX1诱导宫颈癌细胞凋亡和铁死亡有关。Objective:To explore the mechanism of apoptosis and ferroptosis mediated by PAB(Pseudolaric acid B,PAB)in cervical cancer Siha cells.Methods:The cytotoxicity of PAB on Siha was determined using the CCK8 test.Annexin V-FITC and PI double fluorescence labeling were used to detect apoptosis.Fluorescence microscopy was used to assess nuclear morphology and cell death after labeling with PI and Hochest.The cell cycle and intracellular reactive oxygen species(ROS)levels were determined using flow cytometry.Inverted fluorescence microscopy with BODIPY^(TM)581/591 C11 labeling to detect intracellular lipid peroxidation products.Fluorescence microscopy was used to monitor intracellular mitochondrial membrane potential changes using JC-1 labeling.Western blot analysis revealed the expression of ferroptosis-associated proteins GPX4,SLC7A11,Ferritin,and HMOX1,as well as apoptosis-related proteins PARP,c-IAP1,c-IAP2,XIAP,and Survivin.The mRNA expression levels of ferroptosis-related genes were determined using RT-PCR.Results:PAB dramatically reduced the proliferation of cervical cancer Siha cells,induced apoptosis,and stopped the cell cycle.Second,PAB increased intracellular ROS levels,lipid peroxide buildup,and mitochondrial membrane potential increases.Meanwhile,it increased the expression of HMOX1 protein and mRNA,decreased the expression of GPX4,SLC7A11 protein and mRNA,and decreased the expression of Ferritin,c-IAP1,c-IAP2,XIAP,and Survivin.Conclusion:PAB dramatically increased cell death in cervical cancer Siha cells,and the mechanism of action may be connected to HMOX1 overexpression to trigger apoptosis and ferroptosis.

关 键 词：土荆皮乙酸 铁死亡 宫颈癌 

分 类 号：R737.33[医药卫生—肿瘤]