Esculetin,a natural dihydroxy coumarin derived from the Chinese herbal medicine Cortex Fraxini,has demonstrated significant pharmacological activities,including anticancer properties.Ferroptosis,an iron-dependent form...Esculetin,a natural dihydroxy coumarin derived from the Chinese herbal medicine Cortex Fraxini,has demonstrated significant pharmacological activities,including anticancer properties.Ferroptosis,an iron-dependent form of regulated cell death,has garnered considerable attention due to its lethal effect on tumor cells.However,the exact role of ferroptosis in esculetin-mediated anti-hepatocellular carcinoma(HCC)effects remains poorly understood.This study investigated the impact of esculetin on HCC cells both in vitro and in vivo.The findings indicate that esculetin effectively inhibited the growth of HCC cells.Importantly,esculetin promoted the accumulation of intracellular Fe^(2+),leading to an increase in ROS production through the Fenton reaction.This event subsequently induced lipid peroxidation(LPO)and triggered ferroptosis within the HCC cells.The occurrence of ferroptosis was confirmed by the elevation of malondialdehyde(MDA)levels,the depletion of glutathione peroxidase(GSH-Px)activity,and the disruption of mitochondrial morphology.Notably,the inhibitor of ferroptosis,ferrostatin-1(Fer-1),attenuated the anti-tumor effect of esculetin in HCC cells.Furthermore,the findings revealed that esculetin inhibited the Nrf2-xCT/GPx4 axis signaling in HCC cells.Overexpression of Nrf2 upregulated the expression of downstream SLC7A11 and GPX4,consequently alleviating esculetin-induced ferroptosis.In conclusion,this study suggests that esculetin exerts an anti-HCC effect by inhibiting the activity of the Nrf2-xCT/GPx4 axis,thereby triggering ferroptosis in HCC cells.These findings may contribute to the potential clinical use of esculetin as a candidate for HCC treatment.展开更多
Kirsten rat sarcoma virus(KRAS)is a common oncogene in human cancers.Approximately 40%of the patients diagnosed with colorectal cancer(CRC)have KRAS mutations that exhibit strong resistance to targeted molecular thera...Kirsten rat sarcoma virus(KRAS)is a common oncogene in human cancers.Approximately 40%of the patients diagnosed with colorectal cancer(CRC)have KRAS mutations that exhibit strong resistance to targeted molecular therapy and EGFR antibody treatment.In this study,we present photocatalytic silica nanoparticles(A6-FS/BiVO_(4)DMSNs)for targeted therapy of KRAS mutant CRC with the induction of cascadic ferroptosis events.Dendritic mesoporous silica nanoparticles(DMSNs)were impregnated with photocatalytic BiVO_(4),loaded with ferroptotic agents(benzoyl ferrocene:B and sorafenib:S),and encoded with CD44-targeting A6 peptides.For the targeting design,we observed CD44 overexpression in KRAS mutant CRC cells using CPTAC data analysis.Upon laser irradiation,A6-FS/BiVO_(4)DMSNs generate electron-hole pairs(e^(-)/h^(+)),which produce hydroxyl radical(OH·)and superoxide anions(O_(2)·^(-)).Laser irradiation simultaneously initiates the dissociation of iron(Fe^(2+))from benzoyl ferrocene and the release of sorafenib.This cascade induces ferroptosis in KRAS mutant CRC cells,especially under conditional inhibition of redox-regulating proteins(cystine/glutamate antiporter and glutathione peroxidase 4),and significantly inhibits tumor growth in a KRAS mutant CRC xenograft animal model.展开更多
As one of the key targets of tumor metabolic therapy,glucose dyshomeostasis by disrupting glucose metabolism possesses the potential to reverse therapeutic resistance of a variety of regulated cell deaths(RCDs),but th...As one of the key targets of tumor metabolic therapy,glucose dyshomeostasis by disrupting glucose metabolism possesses the potential to reverse therapeutic resistance of a variety of regulated cell deaths(RCDs),but the functional pathways are not fully revealed and employed.Herein,we demonstrate that the intervention on SLC7A11/GSH/GPX4 antioxidant axis by glucose dyshomeostasis can simultaneously promote disulfidptosis,cuproptosis and ferroptosis,which is verified by employing glucose oxidase(GOx)-modified copper-apigenin(CuAp)network nanoshuttles(CuAp@GOx NSs)in ovarian tumor therapy.Ap and GOx can jointly induce glucose dyshomeostasis respectively by inhibiting glucose transporter 1-mediated glucose uptake upstream,and consuming massive glucose downstream.As a result of glucose dyshomeostasis,the NADPH supplement is downregulated,which further disrupts SLC7A11/GSH/GPX4 antioxidant axis.This simultaneously boosts disulfidptosis by facilitating cystine accumulation,cuproptosis by attenuating GSH-mediated Cu+inactivation,and ferroptosis by downregulating GPX4 expression.Owing to the combination of disulfidptosis,cuproptosis and ferroptosis,CuAp@GOx NSs exhibit good efficacy in treating ovarian tumor model.This work proposes an alternative strategy for tumor therapy based on glucose dyshomeostasis,which mainly targets the RCDs relating to SLC7A11/GSH/GPX4 axis.展开更多
铁是几乎所有生物系统中必不可少的金属。然而,细胞内的铁含量需要严格调节,因为过量的铁会随着ROS的产生而产生破坏性影响,铁死亡是一种新型的由铁依赖性脂质过氧化作用引起的氧化调节细胞死亡。铁死亡涉及遗传、代谢和蛋白质的调节、...铁是几乎所有生物系统中必不可少的金属。然而,细胞内的铁含量需要严格调节,因为过量的铁会随着ROS的产生而产生破坏性影响,铁死亡是一种新型的由铁依赖性脂质过氧化作用引起的氧化调节细胞死亡。铁死亡涉及遗传、代谢和蛋白质的调节、触发和执行机制,在很大程度上与其他形式的受调节细胞死亡不重叠。脂质代谢对于肿瘤的增殖以及侵袭、转染都具有很重要的作用,例如细胞膜层面的高水平膜脂质与肿瘤细胞抵御活性氧(ROS)的损伤密切相关,由于高饱和度的膜脂对于氧化反应的刺激不敏感这为肿瘤细胞无形中提供了保护作用。目前主流的肿瘤化疗采用诱导凋亡的方式杀死或抑制肿瘤细胞。然而,越来越明显地是,肿瘤细胞可能对这些依赖凋亡的抗肿瘤方式表现出内在或获得性抵抗,从而大大增加了治疗失败和治疗后复发的风险。对于肿瘤而言,越来越多研究表明,在肿瘤组织中,铁死亡被显著抑制,导致了肿瘤不受控制地转移以及侵袭,这也为调节靶向肿瘤细胞的死亡对于肿瘤的免疫治疗提供了一种新的治疗方式以及为其他以异常细胞增生的疾病提供新的治疗思路。有趣的是,治疗耐药的癌细胞,特别是那些间充质状态和易于转移的癌细胞,非常容易发生铁死亡。目前许多研究证实:铁死亡与肿瘤存在密切的联系,并且涉及一系列机制,并且铁死亡还可以根除一些凋亡不敏感的肿瘤细胞,激活铁死亡途径可能是克服传统癌症治疗耐药机制的潜在策略。故此铁死亡的相关研究进展对于肿瘤的治疗而言具有重要的意义。Iron is an essential metal in almost all biological systems. However, the intracellular iron content needs to be strictly regulated because excessive iron can have destructive effects along with the generation of reactive oxygen species (ROS). Ferroptosis is a novel type of oxidative regulated cell death caused by iron-dependent lipid peroxidation. Ferroptosis involves the regulatory, triggering and execution mechanisms of genetics, metabolism and proteins, and largely does not overlap with other forms of regulated cell death. Lipid metabolism plays a very important role in the proliferation. High levels of membrane lipids at the cellular membrane level are highly correlated with the ability of tumor cells to avoid damage from reactive oxygen species (ROS). Because membrane lipids with high saturation are insensitive to the stimuli of oxidation reactions, they provide an invisible protective effect for tumor cells. Currently, the mainstream tumor chemotherapy uses the method of inducing apoptosis to kill or inhibit tumor cells. However, it is becoming increasingly evident that tumor cells may exhibit intrinsic or acquired resistance to these apoptosis-dependent anti-tumor approaches, which greatly increases the risk of treatment failure and recurrence after treatment. For tumors, more and more studies have shown that in tumor tissues, ferroptosis is significantly inhibited, leading to uncontrolled metastasis and invasion of tumors. This also provides a new treatment approach for tumor immunotherapy by regulating the death of targeted tumor cells, and offers new treatment ideas for other diseases characterized by abnormal cell proliferation. Interestingly, cancer cells resistant to treatment, especially those in a mesenchymal state and prone to metastasis, are highly susceptible to ferroptosis. Currently, many studies have confirmed that there is a close relationship between ferroptosis and tumors, involving a series of mechanisms. Moreover, ferroptosis can also eradicate some tumor cells that are insensitive to apoptosis. Activating the ferroptosis pathway may be a potential strategy to overcome the resistance mechanisms of traditional cancer treatments. Therefore, the relevant research progress of ferroptosis is of great significance for the treatment of tumors.展开更多
Objective:To investigate the mechanism of induction of ferroptosis by brazilin in breast cancer cells.Methods:Breast cancer 4T1 cells were divided into 6 groups:control,brazilin 1/2 half maximal inhibitory concentrati...Objective:To investigate the mechanism of induction of ferroptosis by brazilin in breast cancer cells.Methods:Breast cancer 4T1 cells were divided into 6 groups:control,brazilin 1/2 half maximal inhibitory concentration(IC_(50)),IC_(50),2×IC_(50),erastin(10μg/mL)and capecitabine(10μg/mL)groups.The effect of brazilin on the proliferation of 4T1 cells was detected by cell counting kit-8 assay,and the treatment dose of brazilin was screened.The effect of brazilin on the mitochondrial morphology of 4T1 cells,and the mitochondrial damage was evaluated under electron microscopy.The levels of Fe+,reactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH)and glutathione peroxidase 4(GPX4)were estimated using various detection kits.The invasion and migration abilities of 4T1 cells were detected by scratch assay and transwell assay.The expressions levels of tumor protein p53,solute carrier family 7 member 11(SLC7A11),GPX4 and acyl-CoA synthetase long-chain family member 4(ACSL4)proteins were quantified by Western blot assay.Results:Compared to the control group,the 10(1/2 IC_(50)),20(IC_(50))and 40(2×IC_(50))μg/mL brazilin,erastin,and capecitabine groups showed a significant decrease in the cell survival rate,invasion and migration abilities,GSH,SLC7A11 and GPX4 protein expression levels,and mitochondrial volume and ridge(P<0.05),and a significant increase in the mitochondria membrane density,Fe2+,ROS and MDA levels,and p53 and ACSL4 protein expression levels(P<0.05).Conclusions:Brazilin actuated ferroptosis in breast cancer cells,and the underlying mechanism is mainly associated with the p53/SLC7A11/GPX4 signaling pathway.展开更多
目的:验证心肌细胞缺氧/复氧(H/R)损伤中是否有铁死亡的发生,以及探讨铁死亡抑制剂Ferrostatin-1(Fer-1)对心肌细胞H/R损伤的作用及其机制。方法:新生1~3 d SD乳鼠,提取原代心肌细胞,随机分为正常对照组(Control)、H/R组和H/R+Fer-1组。...目的:验证心肌细胞缺氧/复氧(H/R)损伤中是否有铁死亡的发生,以及探讨铁死亡抑制剂Ferrostatin-1(Fer-1)对心肌细胞H/R损伤的作用及其机制。方法:新生1~3 d SD乳鼠,提取原代心肌细胞,随机分为正常对照组(Control)、H/R组和H/R+Fer-1组。H/R组细胞培养52 h后,加入4 mmol/L Na 2 S 2 O 4溶液,缺氧1 h后,用含10%小牛血清的DMEM培养液复氧培养3 h。H/R+Fer-1组经Fer-1(2μmol/L)预处理24 h后再进行缺氧复氧处理。各组细胞应用紫外分光光度法检测乳酸脱氢酶(LDH)释放率,CCK-8法检测细胞存活率,黄嘌呤氧化酶法检测超氧化物歧化酶(SOD),化学比色法检测丙二醛(MDA),免疫荧光观测线粒体膜电位、活性氧(ROS)改变,Western blot检测铁死亡关键蛋白ACSL4、GPX4的表达。结果:与control组比较,H/R组细胞活性、SOD释放量和MMP水平均显著降低(P<0.05),LDH、MDA、ROS释放量均显著增加(P<0.05),ACSL4蛋白表达显著升高(P<0.05)、GPX4蛋白表达显著下降(P<0.05)。与H/R组比较,H/R+Fer-1组细胞活性、SOD释放量和MMP水平均显著升高(P<0.05),LDH、MDA、ROS释放量均显著降低(P<0.05),ACSL4蛋白表达显著下降(P<0.05)、GPX4蛋白表达显著升高(P<0.05)。结论:心肌细胞H/R损伤中有铁死亡的发生,Fer-1可通过调控ACSL4和GPX4抑制细胞内ROS的产生,从而减轻铁死亡引起的原代心肌细胞缺氧复氧损伤。展开更多
背景:铁死亡介导的缺血再灌注损伤对压疮的发生发展起重要作用,可能存在压疮相关的铁死亡生物标志物,但其机制尚未阐明。目的:通过生物信息学手段探讨压疮的分子机制,寻找压疮过程中铁死亡相关差异基因,为其临床治疗提供新的视角。方法...背景:铁死亡介导的缺血再灌注损伤对压疮的发生发展起重要作用,可能存在压疮相关的铁死亡生物标志物,但其机制尚未阐明。目的:通过生物信息学手段探讨压疮的分子机制,寻找压疮过程中铁死亡相关差异基因,为其临床治疗提供新的视角。方法:使用GEO数据库和FerrDb数据库下载数据集并进行预处理。对单细胞转录组测序数据进行聚类和占比分析、代谢活性和拟时序分析、细胞通讯分析、铁死亡基因集细胞识别和富集分析,确定铁死亡差异基因,并通过动物实验进一步验证。将20只SD大鼠随机分为正常组和模型组,每组10只。正常组大鼠不进行任何处理,模型组大鼠采用缺血再灌注循环周期模式制备压疮大鼠模型,采用荧光定量PCR和免疫印迹检测压疮大鼠创面组织内差异表达基因和蛋白的变化。结果与结论:(1)单细胞转录组测序数据聚类划分为6种细胞类型,压疮组2型和3型角质形成细胞占比较高。(2)不同细胞亚群之间具有明显的代谢异质性和演变轨迹。(3)2型和3型角质形成细胞在细胞通讯中作用最强,2型角质形成细胞配体-受体强度最佳。(4)2型角质形成细胞铁死亡得分较高,有显著上调或下调的差异基因,并得到27个GO富集条目、20个KEGG富集条目和24个铁死亡相关差异基因,以谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)和长链酯酰辅酶A合成酶4(acyl-CoA synthetase long chain family member 4,ACSL4)为主。(5)动物实验验证了与正常组相比,模型组大鼠铁死亡抑制蛋白GPX4表达下调,铁死亡促进蛋白ACSL4表达上调。上述结果证实,压疮组织中存在铁死亡,GPX4和ACSL4为调控压疮组织中铁死亡的重要基因。展开更多
基金supported by the Natural Science Foundations of Fujian Province(Nos.2021J05063 and 2023J01541)a startup grant for High-level Talents of Fujian Medical University(No.XRCZX2021014)。
文摘Esculetin,a natural dihydroxy coumarin derived from the Chinese herbal medicine Cortex Fraxini,has demonstrated significant pharmacological activities,including anticancer properties.Ferroptosis,an iron-dependent form of regulated cell death,has garnered considerable attention due to its lethal effect on tumor cells.However,the exact role of ferroptosis in esculetin-mediated anti-hepatocellular carcinoma(HCC)effects remains poorly understood.This study investigated the impact of esculetin on HCC cells both in vitro and in vivo.The findings indicate that esculetin effectively inhibited the growth of HCC cells.Importantly,esculetin promoted the accumulation of intracellular Fe^(2+),leading to an increase in ROS production through the Fenton reaction.This event subsequently induced lipid peroxidation(LPO)and triggered ferroptosis within the HCC cells.The occurrence of ferroptosis was confirmed by the elevation of malondialdehyde(MDA)levels,the depletion of glutathione peroxidase(GSH-Px)activity,and the disruption of mitochondrial morphology.Notably,the inhibitor of ferroptosis,ferrostatin-1(Fer-1),attenuated the anti-tumor effect of esculetin in HCC cells.Furthermore,the findings revealed that esculetin inhibited the Nrf2-xCT/GPx4 axis signaling in HCC cells.Overexpression of Nrf2 upregulated the expression of downstream SLC7A11 and GPX4,consequently alleviating esculetin-induced ferroptosis.In conclusion,this study suggests that esculetin exerts an anti-HCC effect by inhibiting the activity of the Nrf2-xCT/GPx4 axis,thereby triggering ferroptosis in HCC cells.These findings may contribute to the potential clinical use of esculetin as a candidate for HCC treatment.
文摘目的探究糖尿病肾病(diabetic kidney disease,DKD)血瘀证大鼠肾损害与肾脏铁死亡的潜在机制。方法将50只SPF级雄性SD大鼠分为对照组、DKD组、DKD血瘀证组。采用腹腔注射链脲佐菌素的方法复制DKD大鼠模型,采用尾静脉注射右旋糖酐的方法复制DKD血瘀证模型。实验过程中观察大鼠血瘀证表现及检测24 h尿蛋白、血清肌酐、血尿素氮、血液流变学指标,采用苏木精-伊红染色、Masson染色、PAS染色观察肾脏的组织形态,采用透射电子显微镜观察铁死亡典型细胞的线粒体变化;采用免疫组织化学法、Western blot法检测肾组织铁死亡相关蛋白[长链酯酰辅酶A合成酶4(Acyl-CoA synthetase long chain family member 4,ACSL4)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)]及肾脏纤维化指标[纤维连接蛋白(fibronectin,FN)、Ⅳ型胶原蛋白(typeⅣcollagen,Col-Ⅳ)]的表达水平。结果与对照组比较,DKD组大鼠肾脏病理变化加重,线粒体损伤明显,24 h尿蛋白含量,血清肌酐、血尿素氮水平及全血黏度、血浆黏度明显升高(P<0.05),肾脏ACSL4、FN和Col-Ⅳ及其mRNA表达水平明显升高(P<0.05),GPX4蛋白及其mRNA表达水平明显降低(P<0.05)。DKD血瘀证大鼠出现唇色黯淡、眼球黯红、耳廓紫红、舌下脉络紫黯等血瘀证表现,24 h尿蛋白和血清肌酐、血尿素氮水平明显高于DKD组,光学显微镜下可见肾脏出现明显的系膜基质增生、肾小球萎缩、肾间质胶原沉积和纤维化,电子显微镜下可见肾组织细胞线粒体损伤明显,嵴基本断裂,ACSL4、FN、Col-Ⅳ及其mRNA表达水平较DKD组明显上升(P<0.05),GPX4蛋白及其mRNA表达水平较DKD组明显下降(P<0.05)。结论DKD血瘀证大鼠肾脏损害更加严重,其机制可能与GPX4、ACSL4介导的铁死亡有关。
基金supported by the Basic Science Research Program and Korea Health Technology R&D Project of the National Research Foundation(NRF)funded by the Korean government(Ministry of Education,Ministry of Science and ICT)(Grant Nos.:RS-2023-00208587,RS-2024-00440714,and 2018R1A6A1A 03025523,Republic of Korea).
文摘Kirsten rat sarcoma virus(KRAS)is a common oncogene in human cancers.Approximately 40%of the patients diagnosed with colorectal cancer(CRC)have KRAS mutations that exhibit strong resistance to targeted molecular therapy and EGFR antibody treatment.In this study,we present photocatalytic silica nanoparticles(A6-FS/BiVO_(4)DMSNs)for targeted therapy of KRAS mutant CRC with the induction of cascadic ferroptosis events.Dendritic mesoporous silica nanoparticles(DMSNs)were impregnated with photocatalytic BiVO_(4),loaded with ferroptotic agents(benzoyl ferrocene:B and sorafenib:S),and encoded with CD44-targeting A6 peptides.For the targeting design,we observed CD44 overexpression in KRAS mutant CRC cells using CPTAC data analysis.Upon laser irradiation,A6-FS/BiVO_(4)DMSNs generate electron-hole pairs(e^(-)/h^(+)),which produce hydroxyl radical(OH·)and superoxide anions(O_(2)·^(-)).Laser irradiation simultaneously initiates the dissociation of iron(Fe^(2+))from benzoyl ferrocene and the release of sorafenib.This cascade induces ferroptosis in KRAS mutant CRC cells,especially under conditional inhibition of redox-regulating proteins(cystine/glutamate antiporter and glutathione peroxidase 4),and significantly inhibits tumor growth in a KRAS mutant CRC xenograft animal model.
基金supported by the National Natural Science Foundation of China(Grant Number 82373399,52303085)the Science and Tech-nology Development Program of Jilin Province(Grant Number 202302001)Special Project from MOST of China.Thanks to Min-gjie Chen(Shanghai NewCore Biotechnology Co.,Ltd.)for providing data analysis and visualization support.
文摘As one of the key targets of tumor metabolic therapy,glucose dyshomeostasis by disrupting glucose metabolism possesses the potential to reverse therapeutic resistance of a variety of regulated cell deaths(RCDs),but the functional pathways are not fully revealed and employed.Herein,we demonstrate that the intervention on SLC7A11/GSH/GPX4 antioxidant axis by glucose dyshomeostasis can simultaneously promote disulfidptosis,cuproptosis and ferroptosis,which is verified by employing glucose oxidase(GOx)-modified copper-apigenin(CuAp)network nanoshuttles(CuAp@GOx NSs)in ovarian tumor therapy.Ap and GOx can jointly induce glucose dyshomeostasis respectively by inhibiting glucose transporter 1-mediated glucose uptake upstream,and consuming massive glucose downstream.As a result of glucose dyshomeostasis,the NADPH supplement is downregulated,which further disrupts SLC7A11/GSH/GPX4 antioxidant axis.This simultaneously boosts disulfidptosis by facilitating cystine accumulation,cuproptosis by attenuating GSH-mediated Cu+inactivation,and ferroptosis by downregulating GPX4 expression.Owing to the combination of disulfidptosis,cuproptosis and ferroptosis,CuAp@GOx NSs exhibit good efficacy in treating ovarian tumor model.This work proposes an alternative strategy for tumor therapy based on glucose dyshomeostasis,which mainly targets the RCDs relating to SLC7A11/GSH/GPX4 axis.
文摘铁是几乎所有生物系统中必不可少的金属。然而,细胞内的铁含量需要严格调节,因为过量的铁会随着ROS的产生而产生破坏性影响,铁死亡是一种新型的由铁依赖性脂质过氧化作用引起的氧化调节细胞死亡。铁死亡涉及遗传、代谢和蛋白质的调节、触发和执行机制,在很大程度上与其他形式的受调节细胞死亡不重叠。脂质代谢对于肿瘤的增殖以及侵袭、转染都具有很重要的作用,例如细胞膜层面的高水平膜脂质与肿瘤细胞抵御活性氧(ROS)的损伤密切相关,由于高饱和度的膜脂对于氧化反应的刺激不敏感这为肿瘤细胞无形中提供了保护作用。目前主流的肿瘤化疗采用诱导凋亡的方式杀死或抑制肿瘤细胞。然而,越来越明显地是,肿瘤细胞可能对这些依赖凋亡的抗肿瘤方式表现出内在或获得性抵抗,从而大大增加了治疗失败和治疗后复发的风险。对于肿瘤而言,越来越多研究表明,在肿瘤组织中,铁死亡被显著抑制,导致了肿瘤不受控制地转移以及侵袭,这也为调节靶向肿瘤细胞的死亡对于肿瘤的免疫治疗提供了一种新的治疗方式以及为其他以异常细胞增生的疾病提供新的治疗思路。有趣的是,治疗耐药的癌细胞,特别是那些间充质状态和易于转移的癌细胞,非常容易发生铁死亡。目前许多研究证实:铁死亡与肿瘤存在密切的联系,并且涉及一系列机制,并且铁死亡还可以根除一些凋亡不敏感的肿瘤细胞,激活铁死亡途径可能是克服传统癌症治疗耐药机制的潜在策略。故此铁死亡的相关研究进展对于肿瘤的治疗而言具有重要的意义。Iron is an essential metal in almost all biological systems. However, the intracellular iron content needs to be strictly regulated because excessive iron can have destructive effects along with the generation of reactive oxygen species (ROS). Ferroptosis is a novel type of oxidative regulated cell death caused by iron-dependent lipid peroxidation. Ferroptosis involves the regulatory, triggering and execution mechanisms of genetics, metabolism and proteins, and largely does not overlap with other forms of regulated cell death. Lipid metabolism plays a very important role in the proliferation. High levels of membrane lipids at the cellular membrane level are highly correlated with the ability of tumor cells to avoid damage from reactive oxygen species (ROS). Because membrane lipids with high saturation are insensitive to the stimuli of oxidation reactions, they provide an invisible protective effect for tumor cells. Currently, the mainstream tumor chemotherapy uses the method of inducing apoptosis to kill or inhibit tumor cells. However, it is becoming increasingly evident that tumor cells may exhibit intrinsic or acquired resistance to these apoptosis-dependent anti-tumor approaches, which greatly increases the risk of treatment failure and recurrence after treatment. For tumors, more and more studies have shown that in tumor tissues, ferroptosis is significantly inhibited, leading to uncontrolled metastasis and invasion of tumors. This also provides a new treatment approach for tumor immunotherapy by regulating the death of targeted tumor cells, and offers new treatment ideas for other diseases characterized by abnormal cell proliferation. Interestingly, cancer cells resistant to treatment, especially those in a mesenchymal state and prone to metastasis, are highly susceptible to ferroptosis. Currently, many studies have confirmed that there is a close relationship between ferroptosis and tumors, involving a series of mechanisms. Moreover, ferroptosis can also eradicate some tumor cells that are insensitive to apoptosis. Activating the ferroptosis pathway may be a potential strategy to overcome the resistance mechanisms of traditional cancer treatments. Therefore, the relevant research progress of ferroptosis is of great significance for the treatment of tumors.
基金Supported by National Administration of Traditional Chinese Medicine Youth Qihuang Scholars Talent Project,the 73rd Batch of Grant from China Postdoctoral Science Foundation(No.2023M731069)Traditional Chinese Medicine Administration Project of Hunan Province(No.B2023086)Health Research Project of Hunan Provincial Health Commission(No.W20243081)。
文摘Objective:To investigate the mechanism of induction of ferroptosis by brazilin in breast cancer cells.Methods:Breast cancer 4T1 cells were divided into 6 groups:control,brazilin 1/2 half maximal inhibitory concentration(IC_(50)),IC_(50),2×IC_(50),erastin(10μg/mL)and capecitabine(10μg/mL)groups.The effect of brazilin on the proliferation of 4T1 cells was detected by cell counting kit-8 assay,and the treatment dose of brazilin was screened.The effect of brazilin on the mitochondrial morphology of 4T1 cells,and the mitochondrial damage was evaluated under electron microscopy.The levels of Fe+,reactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH)and glutathione peroxidase 4(GPX4)were estimated using various detection kits.The invasion and migration abilities of 4T1 cells were detected by scratch assay and transwell assay.The expressions levels of tumor protein p53,solute carrier family 7 member 11(SLC7A11),GPX4 and acyl-CoA synthetase long-chain family member 4(ACSL4)proteins were quantified by Western blot assay.Results:Compared to the control group,the 10(1/2 IC_(50)),20(IC_(50))and 40(2×IC_(50))μg/mL brazilin,erastin,and capecitabine groups showed a significant decrease in the cell survival rate,invasion and migration abilities,GSH,SLC7A11 and GPX4 protein expression levels,and mitochondrial volume and ridge(P<0.05),and a significant increase in the mitochondria membrane density,Fe2+,ROS and MDA levels,and p53 and ACSL4 protein expression levels(P<0.05).Conclusions:Brazilin actuated ferroptosis in breast cancer cells,and the underlying mechanism is mainly associated with the p53/SLC7A11/GPX4 signaling pathway.
文摘目的:验证心肌细胞缺氧/复氧(H/R)损伤中是否有铁死亡的发生,以及探讨铁死亡抑制剂Ferrostatin-1(Fer-1)对心肌细胞H/R损伤的作用及其机制。方法:新生1~3 d SD乳鼠,提取原代心肌细胞,随机分为正常对照组(Control)、H/R组和H/R+Fer-1组。H/R组细胞培养52 h后,加入4 mmol/L Na 2 S 2 O 4溶液,缺氧1 h后,用含10%小牛血清的DMEM培养液复氧培养3 h。H/R+Fer-1组经Fer-1(2μmol/L)预处理24 h后再进行缺氧复氧处理。各组细胞应用紫外分光光度法检测乳酸脱氢酶(LDH)释放率,CCK-8法检测细胞存活率,黄嘌呤氧化酶法检测超氧化物歧化酶(SOD),化学比色法检测丙二醛(MDA),免疫荧光观测线粒体膜电位、活性氧(ROS)改变,Western blot检测铁死亡关键蛋白ACSL4、GPX4的表达。结果:与control组比较,H/R组细胞活性、SOD释放量和MMP水平均显著降低(P<0.05),LDH、MDA、ROS释放量均显著增加(P<0.05),ACSL4蛋白表达显著升高(P<0.05)、GPX4蛋白表达显著下降(P<0.05)。与H/R组比较,H/R+Fer-1组细胞活性、SOD释放量和MMP水平均显著升高(P<0.05),LDH、MDA、ROS释放量均显著降低(P<0.05),ACSL4蛋白表达显著下降(P<0.05)、GPX4蛋白表达显著升高(P<0.05)。结论:心肌细胞H/R损伤中有铁死亡的发生,Fer-1可通过调控ACSL4和GPX4抑制细胞内ROS的产生,从而减轻铁死亡引起的原代心肌细胞缺氧复氧损伤。
文摘背景:铁死亡介导的缺血再灌注损伤对压疮的发生发展起重要作用,可能存在压疮相关的铁死亡生物标志物,但其机制尚未阐明。目的:通过生物信息学手段探讨压疮的分子机制,寻找压疮过程中铁死亡相关差异基因,为其临床治疗提供新的视角。方法:使用GEO数据库和FerrDb数据库下载数据集并进行预处理。对单细胞转录组测序数据进行聚类和占比分析、代谢活性和拟时序分析、细胞通讯分析、铁死亡基因集细胞识别和富集分析,确定铁死亡差异基因,并通过动物实验进一步验证。将20只SD大鼠随机分为正常组和模型组,每组10只。正常组大鼠不进行任何处理,模型组大鼠采用缺血再灌注循环周期模式制备压疮大鼠模型,采用荧光定量PCR和免疫印迹检测压疮大鼠创面组织内差异表达基因和蛋白的变化。结果与结论:(1)单细胞转录组测序数据聚类划分为6种细胞类型,压疮组2型和3型角质形成细胞占比较高。(2)不同细胞亚群之间具有明显的代谢异质性和演变轨迹。(3)2型和3型角质形成细胞在细胞通讯中作用最强,2型角质形成细胞配体-受体强度最佳。(4)2型角质形成细胞铁死亡得分较高,有显著上调或下调的差异基因,并得到27个GO富集条目、20个KEGG富集条目和24个铁死亡相关差异基因,以谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)和长链酯酰辅酶A合成酶4(acyl-CoA synthetase long chain family member 4,ACSL4)为主。(5)动物实验验证了与正常组相比,模型组大鼠铁死亡抑制蛋白GPX4表达下调,铁死亡促进蛋白ACSL4表达上调。上述结果证实,压疮组织中存在铁死亡,GPX4和ACSL4为调控压疮组织中铁死亡的重要基因。
