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LncFASA promotes cancer ferroptosis via modulating PRDX1 phase separation

Science China(Life Sciences) 2024年第3期67卷 488-503页

作者： Xiao Fan Fangzhou Liu Xiang Wang Ying Wang Yu Chen Chengyu Shi Xinwan Su Manman Tan Qingfeng Yan Jinrong Peng Jianzhong Shao Yan Xiong Aifu Lin MOE Laboratory of Biosystem Homeostasis and Protection College of Life SciencesZhejiang UniversityHangzhou 310058China Key Laboratoryof Cancer Prevention and Intervention China National Ministry of EducationHangzhou 310009China Cancer Center Zhejiang UniversityHangzhou 310058China Key Laboratory for Cell and Gene Engineering of Zhejiang Province Hangzhou 310058China Department of Central Laboratory the First People's Hospital of HuzhouHuzhou 313000China Key Laboratory of Integrated Oncology and Intelligent Medicine of Zhejiang Province Affiliated Hangzhou First People's HospitalZhejiang University School of MedicineHangzhou 310006China MOE Key Laboratory for Molecular Animal Nutrition College of Animal SciencesZhejiang UniversityHangzhou 310058China Department of Orthopedic Surgery the Second Affiliated HospitalSchool of MedicineZhejiang UniversityHangzhou 310000China Breast Center of the First Affiliated Hospital School of MedicineZhejiang UniversityHangzhou 310003China International School of Medicine International Institutes of MedicineThe 4th Affiliated Hospital of Zhejiang University School of MedicineYiwu 322000China

Ferroptosis, a unique type of non-apoptotic cell death resulting from iron-dependent lipid peroxidation, has a potential physiological function in tumor suppression, but its underlying mechanisms have not been fully elucidated. Here, we report that the long non-coding RNA(lncRNA) LncFASA increases the susceptibility of triple-negative breast cancer(TNBC) to ferroptosis. As a tumor suppressor, LncFASA drives the formation of droplets containing peroxiredoxin1(PRDX1), a member of the peroxidase family, resulting in the accumulation of lipid peroxidation via the SLC7A11-GPX4 axis. Mechanistically, LncFASA directly binds to the Ahpc-TSA domain of PRDX1, inhibiting its peroxidase activity by driving liquid-liquid phase separation, which disrupts intracellular ROS homeostasis. Notably, high LncFASA expression indicates favorable overall survival in individuals with breast cancer, and LncFASA impairs the growth of breast xenograft tumors by modulating ferroptosis. Together, our findings illustrate the crucial role of this lncRNA in ferroptosis-mediated cancer development and provide new insights into therapeutic strategies for breast cancer.

关键词： long non-coding RNA peroxiredoxin liquid-liquid phase separation ferroptosis breast cancer

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Multiple Redox and Non-Redox Interactions Define 2-Cys peroxiredoxin as a Regulatory Hub in the Chloroplast

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Molecular Plant 2009年第6期2卷 1273-1288页

作者： Meenakumari Muthuramalingam Thorsten Seidel Miriam Laxa Susana M. Nunes de Miranda Florian Gartner Elke Stroher Andrea Kandlbinder Karl-Josef Dietz Biochemistry and Physiology of Plants W5-134 Bielefeld University 33501 Bielefeld Germany

In plants, the highly abundant 2-cysteine peroxiredoxin （2-CysPrx） is associated with the chloroplast and involved in protecting photosynthesis. This work addresses the multiple interactions of the 2-CysPrx in the chloroplast, which depend on its redox state. Transcript co-regulation analysis showed a strong linkage to the peptidyl-prolyl-cis/trans isomerase Cyclophilin 20-3 （Cyp20-3） and other components of the photosynthetic apparatus. Co-expression in protoplasts and quantification of fluorescence resonance energy transfer （FRET） efficiency in vivo confirmed protein interactions of 2-CysPrx with Cyp20-3 as well as NADPH-dependent thioredoxin reductase C （NTRC）, while thioredoxin x （Trx-x） did not form complexes that could enable FRET. Likewise, changes in FRET of fluorescently labeled 2-CysPrx in vitro and in vivo proved redox dependent dynamics of 2-CysPrx. Addition of Cyp20-3 to an in vitro peroxidase assay with 2-CysPrx had no significant effect on peroxide reduction. Also, in the presence of NTRC, addition of Cyp20-3 did not further enhance peroxide reduction. In addition, 2-CysPrx functioned as chaperone and inhibited aggregation of citrate synthase during heat treatment. This activity was partly inhibited by Cyp20-3. As a new interaction partner of decameric 2-CysPrx, photosystem Ⅱ could be identified after chloroplast fractionation and in pull-down assays after reconstitution. In summary, the data indicate a dynamic function of plant 2-CysPrx as redox sensor, chaperone, and regulator in the chloroplast with diverse functions beyond its role as thiol peroxidase.

关键词： Oxidative and photo-oxidative stress photosynthesis chloroplast biology Arabidopsis cyclophilin peroxiredoxin thioredoxin.

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特发性弱精子症患者精子中peroxiredoxin I与精浆活性氧的关系

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中南大学学报（医学版） 2014年第8期39卷 842-848页

作者：王海燕刘能辉曾鸿中南大学湘雅医院妇产科生殖医学中心长沙410008

目的:探讨peroxiredoxin I在特发性弱精子症患者精子中的表达情况及其与精浆活性氧(reactive oxygen species,ROS)的关系。方法:选择2012年9月至12月在中南大学湘雅医院生殖医学中心就诊的男性不育患者。实验组为26名特发性弱精子症患者... 详细信息

目的:探讨peroxiredoxin I在特发性弱精子症患者精子中的表达情况及其与精浆活性氧(reactive oxygen species,ROS)的关系。方法:选择2012年9月至12月在中南大学湘雅医院生殖医学中心就诊的男性不育患者。实验组为26名特发性弱精子症患者,选取15名有生育史且精液分析各参数正常的男性为对照组,收集新鲜精液。应用鲁米诺化学发光法检测精浆中ROS水平,蛋白质免疫印迹技术检测peroxiredoxin I在特发性弱精子症患者及对照组精子中表达的差异。结果:1)实验组精浆ROS水平增高,与对照组相比差异有统计学意义(Pperoxiredoxin I的表达显著低于对照组(Pperoxiredoxin I含量与精浆ROS水平也呈负相关(r=–0.474,Pperoxiredoxin I的含量与前向运动精子活动率呈正相关(r=0.779,Pperoxiredoxin I表达下降可能是引起特发性弱精子症发生的重要环节之一。高水平ROS可能是引起特发性弱精子症精子活力下降的主要原因之一。

关键词： peroxiredoxin I 特发性弱精子症活性氧巯基氧化应激

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peroxiredoxin6在胃癌中的表达及其临床意义

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中华医学杂志 2012年第34期92卷 2433-2435页

作者：郭红伟吕有勇朱娅丽杨桂彬北京大学航天临床医学院航天中心医院消化科 100049 北京大学航天临床医学院航天中心医院综合内科 100049 北京大学临床肿瘤学院北京市肿瘤防治研究所分子生物学实验室

目的明确peroxiredoxin6（Prx6）在胃癌中的表达及其临床意义。方法选择1992至2000年北京大学临床肿瘤学院、北京大学第一医院及内蒙古医学院的胃癌病例手术切除后的石蜡包埋组织，其中胃癌86例，癌旁形态学正常组织69例，通过查阅病理... 详细信息

目的明确peroxiredoxin6（Prx6）在胃癌中的表达及其临床意义。方法选择1992至2000年北京大学临床肿瘤学院、北京大学第一医院及内蒙古医学院的胃癌病例手术切除后的石蜡包埋组织，其中胃癌86例，癌旁形态学正常组织69例，通过查阅病理档案获取其临床病理资料；用组织微阵列和免疫组织化学染色方法检测组织中Prx6蛋白表达水平。结果Prx6以细胞质着色为主，胃癌组织中28例（32．6％）表达阳性；癌旁形态学正常组织中65例（94．2％）表达阳性（P〈0．01）。在53例高中分化胃癌组织中，21例（39．6％）Prx6高表达；在33例低分化胃癌组织中，2例（6．1％）Prx6高表达。Prx6在胃癌组织中的表达与患者的年龄、性别、病变部位、浸润深度、临床分期、血管浸润及肝转移均无明显相关性（均P〉0．05）。结论Prx6在胃癌组织中的表达低于癌旁组织，表达强弱与胃癌组织的分化程度有关。

关键词：胃肿瘤活性氧 peroxiredoxin 6

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Effect of peroxiredoxin 6 on total and progressive motility of human spermatozoa after cryopreservation

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BIOCELL 2020年第3期44卷 323-327页

作者： TIECHENG SUN LUYANG CHENG JING MA SHANJIE ZHOU YANDONG ZHANG WENDI ZHOU SHOULONG DENG SHUSONG WANG TIAN LI Reproductive Medical Center Department of Obstetrics and GynecologyPeking University International HospitalBeijing102206China Department of Immunology Basic Medical CollegeChengde Medical UniversityHebei067000China NHC Key Laboratory of Family Planning and Healthy/Key Laboratory of Reproductive Medicine of Hebei Province Shijiazhuang050051China Beijing Key Laboratory for Animal Genetic Improvement National Engineering Laboratory for Animal BreedingKey Laboratory of Animal Genetics and Breeding of the Ministry of AgricultureCollege of Animal Science and TechnologyChina Agricultural UniversityBeijing100193China CAS Key Laboratory of Genome Sciences and Information Beijing Institute of GenomicsChinese Academy of SciencesBeijing100101China Reproductive Medicine Centre Peking University Second Affiliated HospitalBeijing100044China

Sperm cryopreservation is useful in assisted reproductive technology and male fertility preservation.However,freezing and thawing significantly reduces the total and progressive motility of human spermatozoa.In the present study,we explored the effects of peroxiredoxin 6(PRDX6)on total and progressive motility of human spermatozoa after cryopreservation.Semen samples of 20 males with normal parameters were collected and frozen in media supplemented with different concentrations of PRDX6(0 mM,10−5 mM,10−7 mM,and 10−9 mM,respectively).Postthaw total and progressive motility of sperms were measured.The results showed that in comparison with 0 mM,the concentrations of 10−5 mM,10−7 mM,and 10−9 mM of PRDX6 all significantly improved total motility and progressive motility of sperms(p<0.05).The 10−7 mM of PRDX6 showed the best performance.In conclusion,the supplementation of PRDX6 helps to maintain the total and progressive motility of human spermatozoa.

关键词： peroxiredoxin 6 cryopreservation sperm motility progressive motility

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peroxiredoxinⅡ基因在Hep3B细胞中的生物学功能和作用机制

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中华肝脏病杂志 2008年第6期16卷 435-439页

作者：岳海英代智郭坤孙瑞霞曹骥崔杰峰苏建家杨春欧超岳惠芬李瑗刘银坤广西医科大学附属肿瘤医院实验研究部广西南宁530021 复旦大学附属中山医院肝癌研究所上海200032

目的探讨peroxiredoxinⅡ（PrxⅡ）基因在肝癌发生发展中的作用及其机制。方法化学合成两对针对PrxⅡ基因的小干扰RNA，即PrxⅡ1和PrxⅡ2，分别用脂质体Lipofectamine^TM2000介导转入人肝癌细胞系Hep3B。通过实时荧光定量PCR和Western b... 详细信息

目的探讨peroxiredoxinⅡ（PrxⅡ）基因在肝癌发生发展中的作用及其机制。方法化学合成两对针对PrxⅡ基因的小干扰RNA，即PrxⅡ1和PrxⅡ2，分别用脂质体Lipofectamine^TM2000介导转入人肝癌细胞系Hep3B。通过实时荧光定量PCR和Western blot检测确定PrxⅡ在转染细胞中mRNA和蛋白表达水平受到明显抑制后，对PrxⅡ基因沉默的Hep3B细胞分别进行细胞生长、细胞凋亡、细胞集落形成等检测，观察PrxⅡ对Hep3B细胞生物学功能的影响；通过二氯荧光素双乙酸和硫代巴比妥酸试验，检测细胞内过氧化代谢产物活性氧类和丙二醛的水平，探讨PrxⅡ的作用机制。结果与阴性对照组和空白对照组相比，PrxⅡ基因沉默的Hep3B细胞生长受到抑制，生长抑制率在转染96h后达40％以上；细胞凋亡增加；细胞形成集落的能力降低，PrxⅡ-1组和PrxⅡ-2组的细胞集落形成数分别为42.0±2．8和40．5±0．7，明显低于阴性对照组和空白对照组的121．5±2．1和130．0±1．4（P〈0．05）。PrxⅡ基因沉默的Hep3B细胞内活性氧和丙二醛水平明显增加（P〈0．05）。结论PrxⅡ具有促进Hep3B细胞生长的生物学功能，其机制可能与通过抗氧化作用形成有利于肿瘤细胞生长的微环境有关。

关键词：癌肝细胞基因表达 peroxiredoxin Ⅱ RNA干扰

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Celastrol induces ferroptosis in activated HSCs to ameliorate hepatic fibrosis via targeting peroxiredoxins and HO-1

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Acta Pharmaceutica Sinica B 2022年第5期12卷 2300-2314页

作者： Piao Luo Dandan Liu Qian Zhang Fan Yang Yin-Kwan Wong Fei Xia Junzhe Zhang Jiayun Chen Ya Tian Chuanbin Yang Lingyun Dai Han-Ming Shen Jigang Wang Artemisinin Research Center Institute of Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijing 100700China Central People’s Hospital of Zhanjiang Zhanjiang 524037China Department of Urology the Second Clinical Medical CollegeJinan University(Shenzhen People’s Hospital)Shenzhen 518020China Integrated Chinese and Western Medicine Postdoctoral Research Station Jinan UniversityGuangzhou 510632China Faculty of Health Sciences University of MacaoTaipaMacao 999078China Guangdong Provincial Key Laboratory of New Drug Screening School of Pharmaceutical SciencesSouthern Medical UniversityGuangzhou 510515China

Ferroptosis is a form of regulated cell death, characterized by excessive membrane lipid peroxidation in an iron-and ROS-dependent manner. Celastrol, a natural bioactive triterpenoid extracted from Tripterygium wilfordii, shows effective anti-fibrotic and anti-inflammatory activities in multiple hepatic diseases. However, the exact molecular mechanisms of action and the direct protein targets of celastrol in the treatment of liver fibrosis remain largely elusive. Here, we discover that celastrol exerts anti-fibrotic effects via promoting the production of reactive oxygen species(ROS) and inducing ferroptosis in activated hepatic stellate cells(HSCs). By using activity-based protein profiling(ABPP) in combination with bio-orthogonal click chemistry reaction and cellular thermal shift assay(CETSA), we show that celastrol directly binds to peroxiredoxins(PRDXs), including PRDX1, PRDX2, PRDX4 and PRDX6,through the active cysteine sites, and inhibits their anti-oxidant activities. Celastrol also targets to heme oxygenase 1(HO-1) and upregulates its expression in activated-HSCs. Knockdown of PRDX1, PRDX2,PRDX4, PRDX6 or HO-1 in HSCs, to varying extent, elevated cellular ROS levels and induced ferroptosis. Taken together, our findings reveal the direct protein targets and molecular mechanisms via which celastrol ameliorates hepatic fibrosis, thus supporting the further development of celastrol as a promising therapeutic agent for liver fibrosis.

关键词： Celastrol Ferroptosis peroxiredoxin HO-1 Hepatic fibrosis ABPP Anti-oxidant Reactive oxygen species

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peroxiredoxin 1基因沉默对肝癌细胞HepG2和SMMC-7721放射增敏的研究

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西安交通大学学报（医学版） 2011年第5期32卷 569-575页

作者：高麦仓张龙张殿增张军西安交通大学医学院第二附属医院消化科陕西西安710004 西安交通大学医学院第一附属医院肿瘤放疗科陕西西安710061 西安交通大学医学院实验中心陕西西安710061

目的探讨peroxiredoxin（Prx）1基因的小分子干扰RNA（si RNA）对人肝癌细胞系HepG2和SMMC-7721的放射增敏作用及机制。方法通过逆转录PCR方法研究Prx家族（Prx1~6）在人肝癌细胞HepG2和SMMC-7721中的mRNA表达谱;使用si RNA敲低HepG2和S... 详细信息

目的探讨peroxiredoxin（Prx）1基因的小分子干扰RNA（si RNA）对人肝癌细胞系HepG2和SMMC-7721的放射增敏作用及机制。方法通过逆转录PCR方法研究Prx家族（Prx1~6）在人肝癌细胞HepG2和SMMC-7721中的mRNA表达谱;使用si RNA敲低HepG2和SMMC-7721细胞高表达的Prx1亚型为Prx1 si RNA转染组,另设空白对照组、阴性对照组（Prx1si Neg）,分别经不同剂量X射线照射后,克隆形成法检测各组细胞的增殖情况,流式细胞仪测定细胞内活性氧水平、细胞周期及凋亡情况。结果 HepG2和SMMC-7721细胞均高表达Prx1、Prx3和Prx5。与对照组相比,两种细胞Prx1 si RNA转染组在8 Gy X线照射后其剂量生存曲线明显左移,照射12 h后细胞周期出现G2-M阻滞,24 h后细胞凋亡率及1 h后细胞内ROS水平明显升高（P〈0.05）。Prx1 si RNA转染组放射增敏比在1.38~1.45之间。结论小分子干扰RNA对人肝癌HepG2和SMMC-7721细胞有明显的放射增敏作用,其机制可能与细胞周期阻滞及细胞内活性氧水平升高有关。

关键词：肝癌细胞放射敏感性 peroxiredoxin 1 RNA干扰细胞内活性氧细胞周期阻滞放射增敏

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人源噬菌体抗体对peroxiredoxin Ⅰ高表达肺腺癌细胞增殖的抑制作用

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癌症 2009年第10期28卷 1061-1066页

作者：罗弋庞华李淑杰曹辉李少林樊春波重庆医科大学放射医学教研室重庆400016 重庆医科大学附属第一医院核医学科重庆400016

背景与目的:研究表明过氧化物酶peroxiredoxin Ⅰ(Prx Ⅰ)与癌症的发展有密切关系。我们已通过噬菌体展示技术构建了肺腺癌相关的人源单链抗体库。本研究对该库进行筛选,得到抗Prx Ⅰ肺腺癌单链抗体,并检测其对肺腺癌细胞A549增殖的抑... 详细信息

背景与目的:研究表明过氧化物酶peroxiredoxin Ⅰ(Prx Ⅰ)与癌症的发展有密切关系。我们已通过噬菌体展示技术构建了肺腺癌相关的人源单链抗体库。本研究对该库进行筛选,得到抗Prx Ⅰ肺腺癌单链抗体,并检测其对肺腺癌细胞A549增殖的抑制作用。方法:PCR法检测TG1中scFv基因插入率,1%琼脂糖凝胶电泳鉴定Sfi Ⅰ和Not Ⅰ双酶切质粒的结果,以A549细胞及在肺癌中高表达的抗氧化蛋白Prx Ⅰ为靶抗原分别对抗体库进行3轮筛选富集。将阳性克隆用IPTG诱导表达并进行检测。放射性核素计数法测定细胞单链抗体内摄水平,MTT法及流式细胞术检测单链抗体对A549细胞的增殖抑制和凋亡情况,免疫印迹法检测抗体作用A549细胞后Prx Ⅰ的表达水平。结果:scFv基因插入率为77%,双酶切鉴定检测到目的条带。在亲和筛选过程中,肺腺癌单链抗体得到富集,收获率逐轮提高,第6轮为第1轮的180倍。ELISA法检测到在随机选取的10个克隆中,有6个与A549细胞呈阳性反应,阳性率60%。SDS-PAGE及ELISA检测证实得到人源抗PrxI肺腺癌单链抗体。被A549细胞内摄的单链抗体介导了细胞的凋亡以及细胞内Prx Ⅰ蛋白表达水平的下降。结论:从噬菌体抗体库中筛选获得具有较高特异性的抗Prx Ⅰ肺腺癌单链抗体。单链抗体与肺腺癌细胞有特异性亲和力,并能有效抑制其增殖。

关键词：肺腺癌噬菌体抗体库单链抗体 peroxiredoxin I 增殖抑制

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18beta-glycyrrhetinic acid induces ROS-mediated apoptosis to ameliorate hepatic fibrosis by targeting PRDX1/2 in activated HSCs

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Journal of Pharmaceutical Analysis 2022年第4期12卷 570-582页

作者： Qian Zhang Piao Luo Liuhai Zheng Jiayun Chen Junzhe Zhang Huan Tang Dandan Liu Xueling He Qiaoli Shi Liwei Gu Jiahao Li Qiuyan Guo Chuanbin Yang Yin Kwan Wong Fei Xia Jigang Wang Artemisinin Research Center and Institute of Chinese Materia Medica Chinese Academy of Chinese Medical SciencesBeijing100700China Department of Biological Sciences National University of SingaporeSingapore117543Singapore Department of Geriatrics Shenzhen People's Hospital(The Second Clinical Medical CollegeJinan University)Shenzhen518020China

Hepatic stellate cells(HSCs)are essential drivers of fibrogenesis.Inducing activated-HSC apoptosis is a promising strategy for treating hepatic fibrosis.18beta-glycyrrhetinic acid(18b-GA)is a natural compound that exists widely in herbal medicines,such as Glycyrrhiza uralensis Fisch,which is used for treating multiple liver diseases,especially in Asia.In the present study,we demonstrated that 18b-GA decreased hepatic fibrosis by inducing the apoptosis in activated HSCs.18b-GA inhibited the expression of a-smooth muscle actin and collagen type Ⅰ alpha-1.Using a chemoproteomic approach derived from activity-based protein profiling,together with cellular thermal shift assay and surface plasmon resonance,we found that 18b-GA covalently targeted peroxiredoxin 1(PRDX1)and peroxiredoxin 2(PRDX2)proteins via binding to active cysteine residues and thereby inhibited their enzymatic activities.18b-GA induced the elevation of reactive oxygen species(ROS),resulting in the apoptosis of activated HSCs.PRDX1 knockdown also led to ROS-mediated apoptosis in activated HSCs.Collectively,our findings revealed the target proteins and molecular mechanisms of 18b-GA in ameliorating hepatic fibrosis,highlighting the future development of 18b-GA as a novel therapeutic drug for hepatic fibrosis.

关键词： Glycyrrhetinic acid Hepatic fibrosis peroxiredoxin Reactive oxygen species Apoptosis

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