Objective： To test whether the down-regulation of Notchl gene expression by curcumin could inhibit cell growth and induce apoptosis, which may be associated mechanistically with the down-regulation of NF-κB in breas...Objective： To test whether the down-regulation of Notchl gene expression by curcumin could inhibit cell growth and induce apoptosis, which may be associated mechanistically with the down-regulation of NF-κB in breast cancer cells. Methods： Breast cancer cell lines MDA-MB-231 were cultured in vitro and treated with different dosages of curcumin （0, 1.25, 5.0, 20.0μmol/L） for dose-dependent assay and different time （0, 24, 48, 72 h） at the dosage of 5.0μmol/L for time course assay. The changes of the mRNA and protein expression of Notchl and NF-κB were measured by RT-PCR and Western Blot, and MTT assay was used to measure the change of proliferation. Results： The mRNA and protein levels of Notchl and NF-κB were decreased significantly in human breast cancer cell line with the increase of dosage of curcumin（P〈0.05）, and with the extension of time course（P〈0.05）. These changes suggested a dose- and time-dependent manner. The proliferation rate of cells also was significantly inhibited（P〈0.05）. Conclusion： The current results show that the Notch-1 signaling pathway is associated mechanistically with NF-κB activity during curcumin-induced cell growth inhibition and apoptosis of breast cancer cells. These results suggest that the down-regulation of Notch signaling by curcumin may be a novel strategy for the treatment of patients with breast cancer.展开更多
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
Objective To study the mechanistic role of myeloid-specific Notchl knockout inhibiting STING signaling to regulate hepatocyte lipophagy.Methods A mouse model of nonalcoholic steatohepatitis(NASH)was established using ...Objective To study the mechanistic role of myeloid-specific Notchl knockout inhibiting STING signaling to regulate hepatocyte lipophagy.Methods A mouse model of nonalcoholic steatohepatitis(NASH)was established using a high-fat diet(HFD)and mouse bone marrow-derived macrophages(BMMs).Primary hepatocytes were isolated to construct a co-culture system.展开更多
Congenital heart disease(CHD)is the mnost comman birth defect,with 34%of cases attrib utedto genetic variants.NOTCH1,a multi-domain transmembrane protein,regulates heart developmert bycontrolling the differantiation a...Congenital heart disease(CHD)is the mnost comman birth defect,with 34%of cases attrib utedto genetic variants.NOTCH1,a multi-domain transmembrane protein,regulates heart developmert bycontrolling the differantiation and migration of myocardial mesoderm cells,and different variants are presentin differnt types of CHD.In this review,we aim to provide a detailed description of NOTCH1 structuraldomains and their functions,highlighting NOTCH1 variants in CHD and the molecular mechanisms throughwhich they contribute to CHD occurrence,NOTCH1 has two main domains,the NOTCH extracellulardomain(NBCD)and the NOTCH intracellular domain(NICD).NECD facilitates ligand binding and NICDformation,while the NICD functions as a transcrip tion factor,forming complexes with co-factors in thenucleus to initiate gene transcription.Amnong the NOTCH1 variants associated with CHD occurrence,most are loss-of-function variants.Moreover,most of the variants are located in theEGF-like domain.Themolecular mechanism behind the NOTCH1 variant-associated CHD occurrence appears to be either due to aloss-of-function or missense variant.In the loss-of-function mutations,NOTCH1 haploinsufficiency is notedand directly reduces theNICD production,causing CHD ocaurrence.In the less common case of missensevariant,only a mild NOTCH1 malfuncticn is observed,but insufficient to directly lead to CHD occurrence.However,when a missense variant is combined with a risk factor,such as exposure to an environmentaltoxin,the cumulative effect can lead to CHD.Understanding the genetic and molecular mechanisms linkingNOTCH1 variants to CHD is crucial for improving clinical management and patient quality of life.展开更多
文摘Objective： To test whether the down-regulation of Notchl gene expression by curcumin could inhibit cell growth and induce apoptosis, which may be associated mechanistically with the down-regulation of NF-κB in breast cancer cells. Methods： Breast cancer cell lines MDA-MB-231 were cultured in vitro and treated with different dosages of curcumin （0, 1.25, 5.0, 20.0μmol/L） for dose-dependent assay and different time （0, 24, 48, 72 h） at the dosage of 5.0μmol/L for time course assay. The changes of the mRNA and protein expression of Notchl and NF-κB were measured by RT-PCR and Western Blot, and MTT assay was used to measure the change of proliferation. Results： The mRNA and protein levels of Notchl and NF-κB were decreased significantly in human breast cancer cell line with the increase of dosage of curcumin（P〈0.05）, and with the extension of time course（P〈0.05）. These changes suggested a dose- and time-dependent manner. The proliferation rate of cells also was significantly inhibited（P〈0.05）. Conclusion： The current results show that the Notch-1 signaling pathway is associated mechanistically with NF-κB activity during curcumin-induced cell growth inhibition and apoptosis of breast cancer cells. These results suggest that the down-regulation of Notch signaling by curcumin may be a novel strategy for the treatment of patients with breast cancer.
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Objective To study the mechanistic role of myeloid-specific Notchl knockout inhibiting STING signaling to regulate hepatocyte lipophagy.Methods A mouse model of nonalcoholic steatohepatitis(NASH)was established using a high-fat diet(HFD)and mouse bone marrow-derived macrophages(BMMs).Primary hepatocytes were isolated to construct a co-culture system.
基金the National Natural Science Foundation of China,GrantNos.82100321 and 82370353.
文摘Congenital heart disease(CHD)is the mnost comman birth defect,with 34%of cases attrib utedto genetic variants.NOTCH1,a multi-domain transmembrane protein,regulates heart developmert bycontrolling the differantiation and migration of myocardial mesoderm cells,and different variants are presentin differnt types of CHD.In this review,we aim to provide a detailed description of NOTCH1 structuraldomains and their functions,highlighting NOTCH1 variants in CHD and the molecular mechanisms throughwhich they contribute to CHD occurrence,NOTCH1 has two main domains,the NOTCH extracellulardomain(NBCD)and the NOTCH intracellular domain(NICD).NECD facilitates ligand binding and NICDformation,while the NICD functions as a transcrip tion factor,forming complexes with co-factors in thenucleus to initiate gene transcription.Amnong the NOTCH1 variants associated with CHD occurrence,most are loss-of-function variants.Moreover,most of the variants are located in theEGF-like domain.Themolecular mechanism behind the NOTCH1 variant-associated CHD occurrence appears to be either due to aloss-of-function or missense variant.In the loss-of-function mutations,NOTCH1 haploinsufficiency is notedand directly reduces theNICD production,causing CHD ocaurrence.In the less common case of missensevariant,only a mild NOTCH1 malfuncticn is observed,but insufficient to directly lead to CHD occurrence.However,when a missense variant is combined with a risk factor,such as exposure to an environmentaltoxin,the cumulative effect can lead to CHD.Understanding the genetic and molecular mechanisms linkingNOTCH1 variants to CHD is crucial for improving clinical management and patient quality of life.
