Objective:Migrasomes,an emerging class of migration-facilitating membranous extracellular vesicles,remain largely uncharted in the intricate landscape of tumor metastasis.This study aimed to illuminate the roles and m...Objective:Migrasomes,an emerging class of migration-facilitating membranous extracellular vesicles,remain largely uncharted in the intricate landscape of tumor metastasis.This study aimed to illuminate the roles and mechanisms underlying cancer cell-derived migrasomes in breast cancer brain metastasis(BCBM).Methods:Migrasomes were isolated and purified from BCBM cells(231-BR)and non-specific organotropic parental counterparts(MDA-MB-231),specifically designated as Mig-BCBM and Mig-BC,respectively.The role of Mig-BCBM in BCBM was investigated using an in vitro endothelial cell layer permeability model and a BCBM mouse model.The regulatory mechanism underlying Mig-BCBM was assessed using RT-qPCR,western blotting,immunofluorescence,ex vivo fluorescence imaging,and a series of rescue experiments.Results:Mig-BCBM potently augmented the permeability of vascular endothelial layers,which facilitated the efficient migration of 231-BR cells across endothelial barriers in vitro.The administration of Mig-BCBM significantly disrupted the blood-brain barrier(BBB)and accelerated BCBM progression in vivo,as evidenced in mouse models,compared to the Mig-BC and control groups.Mechanistically,Mig-BCBM harbored ATF6,a critical transducer of endoplasmic reticulum(ER)stress.Upon internalization into hCMEC/D3 cells,ATF6 elicited robust ER stress responses,culminating in downregulation of ZO-1 and VE-cadherin.Digital PCR analysis disclosed significant upregulation of ATF6 in serum migrasomes derived from BCBM patients compared to migrasomes from breast cancer patients and healthy individuals.Conclusions:This study uncovered a pivotal role of cancer cell-derived in BCBM by harnessing ATF6-mediated ER stress to disrupt the BBB and promote metastasis,suggesting novel diagnostic and therapeutic strategies targeting migrasomes and migrasome cargo.展开更多
目的研究高血压伴高同型半胱氨酸血症(HHcy)大鼠心肌内质网应激(ERS)时,激活作用转录因子6(ATF6)和C/EBP同源蛋白(CHOP)的表达与左室肥厚的关系及依叶片对其干预效果。方法60只雄性SD大鼠行腹主动脉缩窄术(AAC),术后2周选收缩压(SBP)≥1...目的研究高血压伴高同型半胱氨酸血症(HHcy)大鼠心肌内质网应激(ERS)时,激活作用转录因子6(ATF6)和C/EBP同源蛋白(CHOP)的表达与左室肥厚的关系及依叶片对其干预效果。方法60只雄性SD大鼠行腹主动脉缩窄术(AAC),术后2周选收缩压(SBP)≥140 mm Hg(1 mm Hg=0.133k Pa)大鼠45只随机等分为3组。AAC组普通饲料喂养并双蒸水灌胃;对照组2.5%蛋氨酸饲料喂养并双蒸水灌胃;治疗组2.5%蛋氨酸饲料喂养并依叶片[10 mg/(kg·d)]灌胃。定期测SBP和血同型半胱氨酸(Hcy),查心脏彩超,并用免疫组化和Western blot法观察大鼠心肌ATF6和CHOP的表达。结果术后20周,对照组大鼠SBP,血Hcy,室间隔舒张厚度(IVSD)和左室厚壁舒张厚度(LVPWD),心肌ATF6和CHOP表达均高于AAC组(P<0.05);治疗组大鼠SBP,血Hcy,IVSD和LVPWD,心肌ATF6和CHOP表达均明显低于对照组(P<0.05)。结论高血压伴HHcy大鼠心肌ATF6及CHOP表达较Hcy正常的高血压大鼠明显增高,以凋亡因子CHOP表达更占优势;依叶片有效降压,降低血Hcy,减轻心肌ERS,减缓心肌肥厚的作用。展开更多
Normal spermatogenic processes require the scrotal temperature to be lower than that of the body as excessive heat affects spermatogenesis in the testes,reduces sperm quality and quantity,and even causes infertility.E...Normal spermatogenic processes require the scrotal temperature to be lower than that of the body as excessive heat affects spermatogenesis in the testes,reduces sperm quality and quantity,and even causes infertility.Endoplasmic reticulum stress(ERS)is a crucial factor in many pathologies.Although several studies have linked ERS to heat stress,researchers have not yet determined which ERS signaling pathways contribute to heat-induced testicular damage.Melatonin activates antioxidant enzymes,scavenges free radicals,and protects the testes from inflammation;however,few studies have reported on the influence of melatonin on heat-induced testicular damage.Using a murine model of testicular hyperthermia,we observed that heat stress causes both ERS and apoptosis in the testes,especially in the spermatocytes.These observations were confirmed using the mouse spermatocyte cell line GC2,where the Atf6 and Perk signaling pathways were activated during heat stress.Knockout of the above genes effectively reduced spermatocyte damage caused by heat stress.Pretreatment with melatonin alleviated heat-induced apoptosis by inhibiting the Atf6 and Perk signaling pathways.This mitigation was dependent on the melatonin receptors.In vivo experiments verified that melatonin treatment relieved heat-induced testicular damage.In conclusion,our results demonstrated that ATF6 and PERK are important mediators for heat-induced apoptosis,which can be prevented by melatonin treatment.Thus,our study highlights melatonin as a potential therapeutic agent in mammals for subfertility/infertility induced by testicular hyperthermia.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.81702884)Natural Science Foundation of Shandong Province(Grant Nos.ZR2022MH272,ZR2020QH216,and ZR2023QH115)Medicine and Health Science and Technology Foundation of Shandong Province(Grant Nos.202402060623 and 202202080721).
文摘Objective:Migrasomes,an emerging class of migration-facilitating membranous extracellular vesicles,remain largely uncharted in the intricate landscape of tumor metastasis.This study aimed to illuminate the roles and mechanisms underlying cancer cell-derived migrasomes in breast cancer brain metastasis(BCBM).Methods:Migrasomes were isolated and purified from BCBM cells(231-BR)and non-specific organotropic parental counterparts(MDA-MB-231),specifically designated as Mig-BCBM and Mig-BC,respectively.The role of Mig-BCBM in BCBM was investigated using an in vitro endothelial cell layer permeability model and a BCBM mouse model.The regulatory mechanism underlying Mig-BCBM was assessed using RT-qPCR,western blotting,immunofluorescence,ex vivo fluorescence imaging,and a series of rescue experiments.Results:Mig-BCBM potently augmented the permeability of vascular endothelial layers,which facilitated the efficient migration of 231-BR cells across endothelial barriers in vitro.The administration of Mig-BCBM significantly disrupted the blood-brain barrier(BBB)and accelerated BCBM progression in vivo,as evidenced in mouse models,compared to the Mig-BC and control groups.Mechanistically,Mig-BCBM harbored ATF6,a critical transducer of endoplasmic reticulum(ER)stress.Upon internalization into hCMEC/D3 cells,ATF6 elicited robust ER stress responses,culminating in downregulation of ZO-1 and VE-cadherin.Digital PCR analysis disclosed significant upregulation of ATF6 in serum migrasomes derived from BCBM patients compared to migrasomes from breast cancer patients and healthy individuals.Conclusions:This study uncovered a pivotal role of cancer cell-derived in BCBM by harnessing ATF6-mediated ER stress to disrupt the BBB and promote metastasis,suggesting novel diagnostic and therapeutic strategies targeting migrasomes and migrasome cargo.
文摘目的研究高血压伴高同型半胱氨酸血症(HHcy)大鼠心肌内质网应激(ERS)时,激活作用转录因子6(ATF6)和C/EBP同源蛋白(CHOP)的表达与左室肥厚的关系及依叶片对其干预效果。方法60只雄性SD大鼠行腹主动脉缩窄术(AAC),术后2周选收缩压(SBP)≥140 mm Hg(1 mm Hg=0.133k Pa)大鼠45只随机等分为3组。AAC组普通饲料喂养并双蒸水灌胃;对照组2.5%蛋氨酸饲料喂养并双蒸水灌胃;治疗组2.5%蛋氨酸饲料喂养并依叶片[10 mg/(kg·d)]灌胃。定期测SBP和血同型半胱氨酸(Hcy),查心脏彩超,并用免疫组化和Western blot法观察大鼠心肌ATF6和CHOP的表达。结果术后20周,对照组大鼠SBP,血Hcy,室间隔舒张厚度(IVSD)和左室厚壁舒张厚度(LVPWD),心肌ATF6和CHOP表达均高于AAC组(P<0.05);治疗组大鼠SBP,血Hcy,IVSD和LVPWD,心肌ATF6和CHOP表达均明显低于对照组(P<0.05)。结论高血压伴HHcy大鼠心肌ATF6及CHOP表达较Hcy正常的高血压大鼠明显增高,以凋亡因子CHOP表达更占优势;依叶片有效降压,降低血Hcy,减轻心肌ERS,减缓心肌肥厚的作用。
基金supported by the National Natural Science Foundation of China(32072815)General Project of the Key R&D Plan of Shaanxi Province(2019NY-091)+1 种基金Program of Shaanxi Province Science and Technology Innovation Team(2019TD-036)Fundamental Research Funds for the Central Universities(2452020157)。
文摘Normal spermatogenic processes require the scrotal temperature to be lower than that of the body as excessive heat affects spermatogenesis in the testes,reduces sperm quality and quantity,and even causes infertility.Endoplasmic reticulum stress(ERS)is a crucial factor in many pathologies.Although several studies have linked ERS to heat stress,researchers have not yet determined which ERS signaling pathways contribute to heat-induced testicular damage.Melatonin activates antioxidant enzymes,scavenges free radicals,and protects the testes from inflammation;however,few studies have reported on the influence of melatonin on heat-induced testicular damage.Using a murine model of testicular hyperthermia,we observed that heat stress causes both ERS and apoptosis in the testes,especially in the spermatocytes.These observations were confirmed using the mouse spermatocyte cell line GC2,where the Atf6 and Perk signaling pathways were activated during heat stress.Knockout of the above genes effectively reduced spermatocyte damage caused by heat stress.Pretreatment with melatonin alleviated heat-induced apoptosis by inhibiting the Atf6 and Perk signaling pathways.This mitigation was dependent on the melatonin receptors.In vivo experiments verified that melatonin treatment relieved heat-induced testicular damage.In conclusion,our results demonstrated that ATF6 and PERK are important mediators for heat-induced apoptosis,which can be prevented by melatonin treatment.Thus,our study highlights melatonin as a potential therapeutic agent in mammals for subfertility/infertility induced by testicular hyperthermia.
