目的:探讨组蛋白变异体macroH2A(mH2A)和Butein共同作用GRP78蛋白通过丝裂原激活蛋白激酶(mitogen-activation protein kinase,MAPK)信号通路调控黑色素瘤的生物学行为。方法:采用免疫组织化学检测黑色素瘤和癌旁正常组织中mH2A和葡萄...目的:探讨组蛋白变异体macroH2A(mH2A)和Butein共同作用GRP78蛋白通过丝裂原激活蛋白激酶(mitogen-activation protein kinase,MAPK)信号通路调控黑色素瘤的生物学行为。方法:采用免疫组织化学检测黑色素瘤和癌旁正常组织中mH2A和葡萄糖调节蛋白78(glucose regulated protein of 78,GRP78)的表达情况;分析临床资料与mH2A和GRP78表达的关系;免疫共沉淀实验检测GRP78和mH2A蛋白的相互作用;Western印迹检测mH2A和GRP78的相关作用及Butein与mH2A之间的作用。Transwell试验检测mH2A的表达对黑色素瘤细胞侵袭能力的影响;划痕试验检测mH2A的表达对黑色素瘤细胞迁移能力的影响;Western印迹检测沉默mH2A后细胞外信号调节激酶1(ERK1)和MAPK/ERK激酶(MEK)蛋白的表达。结果:与癌旁正常组织相比,黑色素瘤组织中mH2A和GRP78表达明显降低(P<0.05);Butein可以增强mH2A的表达水平(P<0.05),mH2A可以调节GRP78蛋白的表达(P<0.05);沉默mH2A促进黑色素瘤A375细胞的侵袭迁移能力,上调MEK和ERK1蛋白的表达。结论:Butein可以增强mH2A靶向GRP78蛋白通过MAPK信号通路抑制黑色素瘤细胞的迁移和侵袭行为。展开更多
葡萄糖调节蛋白78(glucose regulated protein 78kD,GRP78)又称免疫球蛋白重链结合蛋白(immunoglobulin heavy chain binding protein,Bip),是位于内质网上重要的分子伴侣,属热休克蛋白70家族的一员,GRP78分子及其DNA分子序列结构在许...葡萄糖调节蛋白78(glucose regulated protein 78kD,GRP78)又称免疫球蛋白重链结合蛋白(immunoglobulin heavy chain binding protein,Bip),是位于内质网上重要的分子伴侣,属热休克蛋白70家族的一员,GRP78分子及其DNA分子序列结构在许多生物物种中高度保守。GRP78在内质网中参与阻止内质网新生肽聚集、调节内质网钙稳态、抗内质网相关性细胞凋亡,以及启动未折叠蛋白反应等细胞生命过程。GRP78基因启动子上存在内质网应激反应元件(ERSE)和cAMP反应元件(CRE)等特殊的顺式作用元件,特异性转录因子ATF6等与GRP78启动子上顺式作用元件发生动态结合,从而调节GRP78基础性或诱导性转录表达。近年来发现,GRP78与脂肪肝、肿瘤和神经系统等疾病的发生发展密切相关,GRP78生物学功能的研究已经引起生物学家们的广泛重视。展开更多
Studies have confirmed a strong association between activation of the endoplasmic reticulum stress pathway and cerebral ischemia/reperfusion(I/R) injury.In this study,three key proteins in the endoplasmic reticulum st...Studies have confirmed a strong association between activation of the endoplasmic reticulum stress pathway and cerebral ischemia/reperfusion(I/R) injury.In this study,three key proteins in the endoplasmic reticulum stress pathway(glucose-regulated protein 78,caspase-12,and C/EBP homologous protein) were selected to examine the potential mechanism of endoplasmic reticulum stress in the neuroprotective effect of G protein-coupled estrogen receptor.Female Sprague-Dawley rats received ovariectomy(OVX),and then cerebral I/R rat models(OVX+ I/R) were established by middle cerebral artery occlusion.Immediately after I/R,rat models were injected with 100 μg/kg E2(OVX + I/R +E2),or 100 μg/kg G protein-coupled estrogen receptor agonist G1(OVX + I/R + G1) in the lateral ventricle.Longa scoring was used to detect neurobehavioral changes in each group.Infarct volumes were measured by 2,3,5-triphenyltetrazolium chloride staining.Morphological changes in neurons were observed by Nissl staining.Terminal dexynucleotidyl transferase-mediated nick end-labeling staining revealed that compared with the OVX + I/R group,neurological function was remarkably improved,infarct volume was reduced,number of normal Nissl bodies was dramatically increased,and number of apoptotic neurons in the hippocampus was decreased after E2 and G1 intervention.To detect the expression and distribution of endoplasmic reticulum stress-related proteins in the endoplasmic reticulum,caspase-12 distribution and expression were detected by immunofluorescence,and mRNA and protein levels of glucose-regulated protein 78,caspase-12,and C/EBP homologous protein were determined by polymerase chain reaction and western blot assay.The results showed that compared with the OVX+ I/R group,E2 and G1 treatment obviously decreased mRNA and protein expression levels of glucose-regulated protein 78,C/EBP homologous protein,and caspase-12.However,the G protein-coupled estrogen receptor antagonist G15(OVX + I/R + E2 + G15) could eliminate the effect of E2 on cerebral I/R injury展开更多
文摘目的:探讨组蛋白变异体macroH2A(mH2A)和Butein共同作用GRP78蛋白通过丝裂原激活蛋白激酶(mitogen-activation protein kinase,MAPK)信号通路调控黑色素瘤的生物学行为。方法:采用免疫组织化学检测黑色素瘤和癌旁正常组织中mH2A和葡萄糖调节蛋白78(glucose regulated protein of 78,GRP78)的表达情况;分析临床资料与mH2A和GRP78表达的关系;免疫共沉淀实验检测GRP78和mH2A蛋白的相互作用;Western印迹检测mH2A和GRP78的相关作用及Butein与mH2A之间的作用。Transwell试验检测mH2A的表达对黑色素瘤细胞侵袭能力的影响;划痕试验检测mH2A的表达对黑色素瘤细胞迁移能力的影响;Western印迹检测沉默mH2A后细胞外信号调节激酶1(ERK1)和MAPK/ERK激酶(MEK)蛋白的表达。结果:与癌旁正常组织相比,黑色素瘤组织中mH2A和GRP78表达明显降低(P<0.05);Butein可以增强mH2A的表达水平(P<0.05),mH2A可以调节GRP78蛋白的表达(P<0.05);沉默mH2A促进黑色素瘤A375细胞的侵袭迁移能力,上调MEK和ERK1蛋白的表达。结论:Butein可以增强mH2A靶向GRP78蛋白通过MAPK信号通路抑制黑色素瘤细胞的迁移和侵袭行为。
文摘葡萄糖调节蛋白78(glucose regulated protein 78kD,GRP78)又称免疫球蛋白重链结合蛋白(immunoglobulin heavy chain binding protein,Bip),是位于内质网上重要的分子伴侣,属热休克蛋白70家族的一员,GRP78分子及其DNA分子序列结构在许多生物物种中高度保守。GRP78在内质网中参与阻止内质网新生肽聚集、调节内质网钙稳态、抗内质网相关性细胞凋亡,以及启动未折叠蛋白反应等细胞生命过程。GRP78基因启动子上存在内质网应激反应元件(ERSE)和cAMP反应元件(CRE)等特殊的顺式作用元件,特异性转录因子ATF6等与GRP78启动子上顺式作用元件发生动态结合,从而调节GRP78基础性或诱导性转录表达。近年来发现,GRP78与脂肪肝、肿瘤和神经系统等疾病的发生发展密切相关,GRP78生物学功能的研究已经引起生物学家们的广泛重视。
基金supported by the National Natural Science Foundation of China,No.81560175,81260159(both to LL)
文摘Studies have confirmed a strong association between activation of the endoplasmic reticulum stress pathway and cerebral ischemia/reperfusion(I/R) injury.In this study,three key proteins in the endoplasmic reticulum stress pathway(glucose-regulated protein 78,caspase-12,and C/EBP homologous protein) were selected to examine the potential mechanism of endoplasmic reticulum stress in the neuroprotective effect of G protein-coupled estrogen receptor.Female Sprague-Dawley rats received ovariectomy(OVX),and then cerebral I/R rat models(OVX+ I/R) were established by middle cerebral artery occlusion.Immediately after I/R,rat models were injected with 100 μg/kg E2(OVX + I/R +E2),or 100 μg/kg G protein-coupled estrogen receptor agonist G1(OVX + I/R + G1) in the lateral ventricle.Longa scoring was used to detect neurobehavioral changes in each group.Infarct volumes were measured by 2,3,5-triphenyltetrazolium chloride staining.Morphological changes in neurons were observed by Nissl staining.Terminal dexynucleotidyl transferase-mediated nick end-labeling staining revealed that compared with the OVX + I/R group,neurological function was remarkably improved,infarct volume was reduced,number of normal Nissl bodies was dramatically increased,and number of apoptotic neurons in the hippocampus was decreased after E2 and G1 intervention.To detect the expression and distribution of endoplasmic reticulum stress-related proteins in the endoplasmic reticulum,caspase-12 distribution and expression were detected by immunofluorescence,and mRNA and protein levels of glucose-regulated protein 78,caspase-12,and C/EBP homologous protein were determined by polymerase chain reaction and western blot assay.The results showed that compared with the OVX+ I/R group,E2 and G1 treatment obviously decreased mRNA and protein expression levels of glucose-regulated protein 78,C/EBP homologous protein,and caspase-12.However,the G protein-coupled estrogen receptor antagonist G15(OVX + I/R + E2 + G15) could eliminate the effect of E2 on cerebral I/R injury
