目前针对恶性胶质瘤的治疗手段十分有限,本研究旨在通过揭示天然产物银杏叶提取物(Ginkgo biloba extract,GBE)的抗肿瘤机制,为恶性胶质瘤的治疗提供新的潜在治疗方案。研究发现,125μg/mL GBE处理A172胶质瘤细胞可显著抑制其活力(EC50=...目前针对恶性胶质瘤的治疗手段十分有限,本研究旨在通过揭示天然产物银杏叶提取物(Ginkgo biloba extract,GBE)的抗肿瘤机制,为恶性胶质瘤的治疗提供新的潜在治疗方案。研究发现,125μg/mL GBE处理A172胶质瘤细胞可显著抑制其活力(EC50=128.6μg/mL),TUNEL及瑞氏染色均观察到典型的细胞凋亡形态。机制上,GBE处理导致线粒体膜电位显著下降(JC-1法检测)并引发内质网扩张(ER-Tracker观察)。Western blot分析证实,GBE激活了线粒体自噬与内质网应激,表现为LC3-Ⅱ/Ⅰ比值升高以及葡萄糖调节蛋白78(glucose-regulated protein 78,GRP78)和C/EBP同源蛋白(C/EBP homologous protein,CHOP)表达上调。与此同时,免疫荧光与Western blot结果显示,Notch同源物1(neurogenic locus notch homolog protein 1,Notch1)/转化生长因子-β1(transforming growth factor-beta1,TGF-β1)/Smad家族成员2/3(Smad family member 2/3,Smad2/3)通路被激活,且Notch1胞内结构域(Notch1 intracellular domain,NICD)发生核转位。双荧光素酶报告实验进一步揭示,Smad2/3可直接促进LC3和CHOP的转录。关键的是,使用Notch1抑制剂(DAPT)或TGF-β1抑制剂(SB431542)进行干预,可部分逆转由GBE诱导的细胞器应激并提升细胞存活率。综上所述,GBE通过激活Notch1/TGF-β1/Smad2/3通路,破坏细胞器稳态,进而诱导A172胶质瘤细胞凋亡。本研究的目的在于系统阐明GBE通过“信号通路-细胞器”互作以发挥抗癌作用的全新分子机制,其意义不仅为GBE作为潜在的胶质瘤治疗药物提供了坚实的实验证据,更揭示了Notch1/TGF-β1/Smad2/3通路可作为未来开发抗胶质瘤新药的重要靶点。展开更多
State Administration for Market Regulation and National Standardization Administration of China have approved the following 14 national guiding technical docoments for standardization.
Ischemia–reperfusion injury is a common pathophysiological mechanism in retinal degeneration.PANoptosis is a newly defined integral form of regulated cell death that combines the key features of pyroptosis,apoptosis,...Ischemia–reperfusion injury is a common pathophysiological mechanism in retinal degeneration.PANoptosis is a newly defined integral form of regulated cell death that combines the key features of pyroptosis,apoptosis,and necroptosis.Oligomerization of mitochondrial voltage-dependent anion channel 1 is an important pathological event in regulating cell death in retinal ischemia–reperfusion injury.However,its role in PANoptosis remains largely unknown.In this study,we demonstrated that voltage-dependent anion channel 1 oligomerization-mediated mitochondrial dysfunction was associated with PANoptosis in retinal ischemia–reperfusion injury.Inhibition of voltage-dependent anion channel 1 oligomerization suppressed mitochondrial dysfunction and PANoptosis in retinal cells subjected to ischemia–reperfusion injury.Mechanistically,mitochondria-derived reactive oxygen species played a central role in the voltagedependent anion channel 1-mediated regulation of PANoptosis by promoting PANoptosome assembly.Moreover,inhibiting voltage-dependent anion channel 1 oligomerization protected against PANoptosis in the retinas of rats subjected to ischemia–reperfusion injury.Overall,our findings reveal the critical role of voltage-dependent anion channel 1 oligomerization in regulating PANoptosis in retinal ischemia–reperfusion injury,highlighting voltage-dependent anion channel 1 as a promising therapeutic target.展开更多
Brassinosteroids(BRs)are essential phytohormones that broadly regulate plant growth,development,and adaptation to biotic and abiotic stresses.In Arabidopsis,apoplastic BR molecules are perceived by a plasma membrane-l...Brassinosteroids(BRs)are essential phytohormones that broadly regulate plant growth,development,and adaptation to biotic and abiotic stresses.In Arabidopsis,apoplastic BR molecules are perceived by a plasma membrane-localized receptor complex comprising the ligand-binding receptor BRI1 and the co-receptor BAK1.While negative regulators of the BR receptor complex,such as BKI1,BIR3,and PUB12/13,have been well characterized,how BRI1 and BAK1 are positively modulated in the BR pathway remains largely unknown.In this study,a genetic screen involving overexpression of RLP genes in the bak1-3 bkk1-1 double mutant reveals that enhanced RLP51 expression partially suppresses the BR-deficient phenotypes of bak1-3 bkk1-1.RLP51 overexpression also partially rescues the weak bri1 mutant allele,bri1-301.Although the rlp51 single mutant exhibits wild-type-like phenotypes,it enhances BR-defective phenotypes in bri1-301 and bak1 serk1 mutants.RLP51 is next found to interact with both BRI1 and BAK1 without affecting BRI1–BAK1 interaction.Critically,co-expression of RLP51 with BRI1 or BAK1 significantly increases BRI1 and BAK1 protein abundances.RLP51 appears to promote protein synthesis rather than stabilize BRI1 and BAK1 proteins.Thus,our study identifies RLP51 as a positive regulator of BR signaling that enhances the protein levels of BRI1 and BAK1.展开更多
目的探究核仁蛋白14(nucleolar protein 14,NOP14)过表达对卵巢癌SKOV3细胞增殖与迁移的影响及机制。方法检测卵巢癌细胞系(SKOV3、A2780、HO-8910、OVCAR)中NOP14的表达水平。SKOV3细胞中转染pcDNA-NOP14质粒以构建NOP14过表达细胞系...目的探究核仁蛋白14(nucleolar protein 14,NOP14)过表达对卵巢癌SKOV3细胞增殖与迁移的影响及机制。方法检测卵巢癌细胞系(SKOV3、A2780、HO-8910、OVCAR)中NOP14的表达水平。SKOV3细胞中转染pcDNA-NOP14质粒以构建NOP14过表达细胞系。细胞克隆形成实验和5-乙炔基-2′-脱氧尿苷(5-Ethynyl-2′-deoxyuridine,EdU)染色检测细胞增殖能力。无血清成球培养分析肿瘤干细胞特性。流式细胞术检测CD133阳性细胞比例。细胞形态学观察及上皮-间质转化(epithelial-mesenchymal transition,EMT)标志蛋白(E-cadherin、N-cadherin、Vimentin)检测评估EMT进程。蛋白质印迹和RT-qPCR检测核受体相互作用蛋白1(nuclear receptor interacting protein 1,NRIP1)及Wnt、β-catenin的表达水平。结果在卵巢癌细胞系中NOP14 mRNA和蛋白表达均显著低于正常卵巢上皮细胞。NOP14过表达后抑制SKOV3细胞增殖、干细胞特性及EMT转化(间质标志物N-cadherin、Vimentin下调,上皮标志物E-cadherin上调),以及NRIP1、Wnt和β-catenin的表达水平。结论NOP14可负向调控NRIP1和Wnt及β-catenin的表达,并抑制卵巢癌SKOV3细胞的增殖、干细胞特性及EMT进程。展开更多
文摘目前针对恶性胶质瘤的治疗手段十分有限,本研究旨在通过揭示天然产物银杏叶提取物(Ginkgo biloba extract,GBE)的抗肿瘤机制,为恶性胶质瘤的治疗提供新的潜在治疗方案。研究发现,125μg/mL GBE处理A172胶质瘤细胞可显著抑制其活力(EC50=128.6μg/mL),TUNEL及瑞氏染色均观察到典型的细胞凋亡形态。机制上,GBE处理导致线粒体膜电位显著下降(JC-1法检测)并引发内质网扩张(ER-Tracker观察)。Western blot分析证实,GBE激活了线粒体自噬与内质网应激,表现为LC3-Ⅱ/Ⅰ比值升高以及葡萄糖调节蛋白78(glucose-regulated protein 78,GRP78)和C/EBP同源蛋白(C/EBP homologous protein,CHOP)表达上调。与此同时,免疫荧光与Western blot结果显示,Notch同源物1(neurogenic locus notch homolog protein 1,Notch1)/转化生长因子-β1(transforming growth factor-beta1,TGF-β1)/Smad家族成员2/3(Smad family member 2/3,Smad2/3)通路被激活,且Notch1胞内结构域(Notch1 intracellular domain,NICD)发生核转位。双荧光素酶报告实验进一步揭示,Smad2/3可直接促进LC3和CHOP的转录。关键的是,使用Notch1抑制剂(DAPT)或TGF-β1抑制剂(SB431542)进行干预,可部分逆转由GBE诱导的细胞器应激并提升细胞存活率。综上所述,GBE通过激活Notch1/TGF-β1/Smad2/3通路,破坏细胞器稳态,进而诱导A172胶质瘤细胞凋亡。本研究的目的在于系统阐明GBE通过“信号通路-细胞器”互作以发挥抗癌作用的全新分子机制,其意义不仅为GBE作为潜在的胶质瘤治疗药物提供了坚实的实验证据,更揭示了Notch1/TGF-β1/Smad2/3通路可作为未来开发抗胶质瘤新药的重要靶点。
文摘State Administration for Market Regulation and National Standardization Administration of China have approved the following 14 national guiding technical docoments for standardization.
基金supported by the National Natural Science Foundation of China,Nos.82172196(to KX),82372507(to KX)the Natural Science Foundation of Hunan Province,China,No.2023JJ40804(to QZ)the Key Laboratory of Emergency and Trauma(Hainan Medical University)of the Ministry of Education,China,No.KLET-202210(to QZ)。
文摘Ischemia–reperfusion injury is a common pathophysiological mechanism in retinal degeneration.PANoptosis is a newly defined integral form of regulated cell death that combines the key features of pyroptosis,apoptosis,and necroptosis.Oligomerization of mitochondrial voltage-dependent anion channel 1 is an important pathological event in regulating cell death in retinal ischemia–reperfusion injury.However,its role in PANoptosis remains largely unknown.In this study,we demonstrated that voltage-dependent anion channel 1 oligomerization-mediated mitochondrial dysfunction was associated with PANoptosis in retinal ischemia–reperfusion injury.Inhibition of voltage-dependent anion channel 1 oligomerization suppressed mitochondrial dysfunction and PANoptosis in retinal cells subjected to ischemia–reperfusion injury.Mechanistically,mitochondria-derived reactive oxygen species played a central role in the voltagedependent anion channel 1-mediated regulation of PANoptosis by promoting PANoptosome assembly.Moreover,inhibiting voltage-dependent anion channel 1 oligomerization protected against PANoptosis in the retinas of rats subjected to ischemia–reperfusion injury.Overall,our findings reveal the critical role of voltage-dependent anion channel 1 oligomerization in regulating PANoptosis in retinal ischemia–reperfusion injury,highlighting voltage-dependent anion channel 1 as a promising therapeutic target.
基金supported by the National Natural Science Foundation of China(32370295 and 32170280 to K.H.)Foundation of Science and Technology of Gansu Province(22ZD6NA049)+3 种基金the Fundamental Research Funds for the Central Universities(lzujbky-2021-kb05 to Y.F.and lzujbky-2023-kb05 to P.L.)the Science and Technology Department of Gansu Province(24JRRA392 to K.H.and 23JRRA1132 to C.X.)China Postdoctoral Science Foundation(2024M751259 and GZB20240285 to B.L.)Science and Technology Program of Gansu Province(25JRRA718 to B.L.).
文摘Brassinosteroids(BRs)are essential phytohormones that broadly regulate plant growth,development,and adaptation to biotic and abiotic stresses.In Arabidopsis,apoplastic BR molecules are perceived by a plasma membrane-localized receptor complex comprising the ligand-binding receptor BRI1 and the co-receptor BAK1.While negative regulators of the BR receptor complex,such as BKI1,BIR3,and PUB12/13,have been well characterized,how BRI1 and BAK1 are positively modulated in the BR pathway remains largely unknown.In this study,a genetic screen involving overexpression of RLP genes in the bak1-3 bkk1-1 double mutant reveals that enhanced RLP51 expression partially suppresses the BR-deficient phenotypes of bak1-3 bkk1-1.RLP51 overexpression also partially rescues the weak bri1 mutant allele,bri1-301.Although the rlp51 single mutant exhibits wild-type-like phenotypes,it enhances BR-defective phenotypes in bri1-301 and bak1 serk1 mutants.RLP51 is next found to interact with both BRI1 and BAK1 without affecting BRI1–BAK1 interaction.Critically,co-expression of RLP51 with BRI1 or BAK1 significantly increases BRI1 and BAK1 protein abundances.RLP51 appears to promote protein synthesis rather than stabilize BRI1 and BAK1 proteins.Thus,our study identifies RLP51 as a positive regulator of BR signaling that enhances the protein levels of BRI1 and BAK1.
文摘目的探究核仁蛋白14(nucleolar protein 14,NOP14)过表达对卵巢癌SKOV3细胞增殖与迁移的影响及机制。方法检测卵巢癌细胞系(SKOV3、A2780、HO-8910、OVCAR)中NOP14的表达水平。SKOV3细胞中转染pcDNA-NOP14质粒以构建NOP14过表达细胞系。细胞克隆形成实验和5-乙炔基-2′-脱氧尿苷(5-Ethynyl-2′-deoxyuridine,EdU)染色检测细胞增殖能力。无血清成球培养分析肿瘤干细胞特性。流式细胞术检测CD133阳性细胞比例。细胞形态学观察及上皮-间质转化(epithelial-mesenchymal transition,EMT)标志蛋白(E-cadherin、N-cadherin、Vimentin)检测评估EMT进程。蛋白质印迹和RT-qPCR检测核受体相互作用蛋白1(nuclear receptor interacting protein 1,NRIP1)及Wnt、β-catenin的表达水平。结果在卵巢癌细胞系中NOP14 mRNA和蛋白表达均显著低于正常卵巢上皮细胞。NOP14过表达后抑制SKOV3细胞增殖、干细胞特性及EMT转化(间质标志物N-cadherin、Vimentin下调,上皮标志物E-cadherin上调),以及NRIP1、Wnt和β-catenin的表达水平。结论NOP14可负向调控NRIP1和Wnt及β-catenin的表达,并抑制卵巢癌SKOV3细胞的增殖、干细胞特性及EMT进程。
