秦巴硒菇提取物FA-2-b-β通过STAT3/GPX4信号通路诱导伯基特淋巴瘤细胞铁死亡  

Agaricus blazei extract FA-2-b-βinduces ferroptosis of Burkitt lymphoma cells through STAT3/GPX4 signaling pathway

作　　者：韦佳 李蓉[2] 王卉妍[2] 席祖军 孙延庆 WEI Jia;LI Rong;WANG Huiyan;XI Zujun;SUN Yanqing(Integrative Chinese and Western Medicine of Gansu University of Chinese Medicine,Lanzhou 730030,China;The First Clinical Medical of Gansu University of Chinese Medicine,Lanzhou 730030,China;Department of Hematology,Gansu Provincial People's Hospital,Lanzhou 730030,China)

机构地区：[1]甘肃中医药大学中西医结合学院,甘肃兰州730030 [2]甘肃中医药大学第一临床医学院,甘肃兰州730030 [3]甘肃省人民医院血液内科,甘肃兰州730030

出　　处：《中国病理生理杂志》2025年第3期453-462,共10页Chinese Journal of Pathophysiology

基　　金：甘肃省科技厅自然科学基金资助项目(No.22JR11RA254);甘肃省人民医院优秀硕/博士生培育计划项目(No.ZX-62000001-2022-678);甘肃省教育厅优秀研究生“创新之星”项目(No.2023CXZX-757)。

摘　　要：目的:探索秦巴硒菇提取物FA-2-b-β诱导伯基特淋巴瘤细胞铁死亡的作用及机制。方法:伯基特淋巴瘤Raji和CA46细胞用FA-2-b-β单独处理,分为对照组、低浓度(2.7 g/L和2.1 g/L)组、中浓度(3.6 g/L和2.8 g/L)组和高浓度(4.5 g/L和3.5 g/L)组;FA-2-b-β与铁死亡抑制剂ferrostatin-1(Fer-1)联合处理,分为对照组、低浓度(2.7 g/L和2.1 g/L)组、中浓度(3.6 g/L和2.8 g/L)组、高浓度(4.5 g/L和3.5 g/L)组和高浓度FA-2-b-β+Fer-1组;高浓度FA-2-b-β联合Fer-1及信号传导及转录激活因子3(signal transducer and activator of transcription 3,STAT3)抑制剂Stattic处理,分为对照组、FA-2-b-β组、FA-2-b-β+Stattic组和FA-2-b-β+Fer-1组。采用CCK-8法测定各组细胞活力,计算FA-2-b-β对Raji和CA46细胞的半数抑制浓度(half-maximal inhibitory concentration,IC50);流式细胞术测定细胞凋亡、线粒体膜电位、细胞周期和活性氧(reactive oxygen species,ROS)水平;采用生化试剂盒检测细胞内Fe2+、丙二醛(malondialdehyde,MDA)和谷胱甘肽(glutathione,GSH)水平;RT-qPCR和Western blot实验测定铁死亡相关分子的mRNA和蛋白表达水平。结果:不同浓度FA-2-b-β作用于Raji和CA46细胞后,与对照组相比,细胞凋亡增加,G0/G1期延长,线粒体膜电位降低(P<0.05);Fe2+、ROS和MDA水平显著升高,GSH水平显著下降(P<0.05);STAT3和谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)的mRNA和蛋白表达下调,p-STAT3蛋白水平降低,前列腺素过氧化物合酶2(prostaglandin-endoperoxide synthase 2,PTSG2)和转铁蛋白受体1(transferrin receptor protein 1,TfR1)的mRNA和蛋白表达上调(P<0.05),溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)的mRNA和蛋白表达水平无显著变化。联合STAT3抑制剂Stattic处理进一步加重上述变化,而铁死亡抑制剂Fer-1则逆转上述指标。结论:秦巴硒菇提取物FA-2-b-β可诱导伯基特淋巴瘤细胞铁死亡,其机制可能与其抑制STAT3/GPX4信号通路有关。AIM:This study aims to investigate the effect of Agaricus blazei extract FA-2-b-βon ferroptosis of Burkitt lymphoma cells and its mechanism.METHODS:Burkitt lymphoma cell lines Raji and CA46 were treated with FA-2-b-βalone and in combination with ferrostatin-1,a ferroptosis inhibitor,or Stattic,a signal transducer and activator of transcription 3(STAT3)inhibitor.Cell viability was assessed using the CCK-8 method,and the half-maximal inhibitory concentration(IC50)of FA-2-b-βwas calculated.Flow cytometry was used to detect apoptosis,cell cycle,mitochondrial membrane potential,and reactive oxygen species(ROS).Additionally,malondialdehyde(MDA)and glutathione(GSH)levels were measured using kits.The mRNA and protein expression levels of ferroptosis-related molecules were determined by RT-qPCR and Western blot.RESULTS:The extract FA-2-b-βat different concentrations significantly inhibited the proliferation of Raji and CA46 cells(P<0.05),promoted their death,regulated cell arrest in G0/G1 phase,and decreased the mitochondrial membrane potential.(2)ROS and MDA levels were significantly increased with different concentrations of the extract FA-2-b-β(P<0.05),while the GSH content was significantly decreased(P<0.05).(3)The protein and mRNA levels of signal transducer and activator of transcription 3(STAT3),p-STAT3,and glutathione peroxidase 4(GPX4)were down-regulated at different concentrations of the extract FA-2-b-β.In addition,prostaglandin-endoperoxide synthase 2(PTSG2)and transferrin receptor protein 1(TfR1)protein and mRNA were up-regulated(P<0.05),while the protein and mRNA levels of solute carrier family 7 member 11(SLC7A11)were not significantly changed.CONCLUSION:The extract FA-2-b-βcan induce ferroptosis in burkitt lymphoma,and the mechanism may be related to the inhibi-tion of STAT3/GPX4 signaling pathway.

关 键 词：秦巴硒菇 FA-2-b-β 伯基特淋巴瘤 铁死亡 STAT3/GPX4信号通路 

分 类 号：R730.2[医药卫生—肿瘤] R966[医药卫生—临床医学] Q291[生物学—细胞生物学]