背景与目的热休克蛋白90AB1(heat shock protein 90 k Da alpha,class B member 1,Hsp90AB1)是ATP依赖的高度保守的分子伴侣,在多种肿瘤细胞中过表达。在肿瘤发生发展的信号传导通路中起着重要作用的一些分子,如表皮生长因子受体(epider...背景与目的热休克蛋白90AB1(heat shock protein 90 k Da alpha,class B member 1,Hsp90AB1)是ATP依赖的高度保守的分子伴侣,在多种肿瘤细胞中过表达。在肿瘤发生发展的信号传导通路中起着重要作用的一些分子,如表皮生长因子受体(epidermal growth factor receptor,EGFR)、人类表皮生长因子受体-2(human epidermal growth factor receptor-2,HER2)等,均是Hsp90AB1的底物蛋白。Hsp90AB1与这些底物蛋白相互作用并参与细胞的多种病理生理过程。本研究通过检测Hsp90AB1在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中的蛋白表达情况以初步探讨其临床意义。方法采用组织微阵列和免疫组织化学染色的方法检测Hsp90AB1在213例NSCLC及相应癌旁正常肺组织中的蛋白表达,并分析Hsp90AB1的表达与NSCLC临床病理参数及患者预后的关系。结果 Hsp90AB1在肺癌组织中的表达水平(阳性率54.0%)高于在正常肺组织中的表达水平(阳性率0.0%,P<0.001)。Hsp90AB1在肺腺癌中的表达阳性率为61.2%,高于肺鳞癌组织37.9%(P=0.002),并且其高表达与肺腺癌患者的不良预后相关(P=0.032)。Hsp90AB1蛋白表达水平与临床分期、淋巴结转移、病理分级等因素无关(P>0.05)。结论 Hsp90AB1在NSCLC组织中高表达,并且其表达水平与肺癌的病理类型及肺腺癌患者总生存期相关。展开更多
As a highly important fish virus,nervous necrosis virus(NNV)has caused severe economic losses to the aquaculture industry worldwide.Autophagy,an evolutionarily conserved intracellular degradation process,is involved i...As a highly important fish virus,nervous necrosis virus(NNV)has caused severe economic losses to the aquaculture industry worldwide.Autophagy,an evolutionarily conserved intracellular degradation process,is involved in the pathogenesis of several viruses.Although NNV can induce autophagy to facilitate infection in grouper fish spleen cells,how it initiates and mediates autophagy pathways during the initial stage of infection is still unclear.Here,we found that red-spotted grouper NNV(RGNNV)induced autophagosome formation in two fish cell lines at 1.5 and 3 h post infection,indicating that autophagy is activated upon entry of RGNNV.Moreover,autophagic detection showed that RGNNV entry induced incomplete autophagy by impairing the fusion of autophagosomes with lysosomes.Further investigation revealed that binding of the RGNNV capsid protein(CP)to the Lateolabrax japonicus heat shock protein HSP90ab1(LjHSP90ab1),a cell surface receptor of RGNNV,contributed to RGNNV invasion-induced autophagy.Finally,we found that CP blocked the interaction of L.japonicus protein kinase B(AKT)with LjHSP90ab1 by competitively binding the NM domain of LjHSP90ab1 to inhibit the AKT-mechanistic target of the rapamycin(MTOR)pathway.This study provides novel insight into the relationship between NNV receptors and autophagy,which may help clarify the pathogenesis of NNV.展开更多
目的:探讨热休克蛋白90ABl(heat shock protein 90 kDa alpha,class B member 1,Hsp90AB1)在肝细胞癌中的表达意义。方法:使用免疫组织化学染色方法检测肝细胞癌组织芯片中Hsp90AB1的表达。运用生物信息学技术分析肝细胞癌相关基因芯片G...目的:探讨热休克蛋白90ABl(heat shock protein 90 kDa alpha,class B member 1,Hsp90AB1)在肝细胞癌中的表达意义。方法:使用免疫组织化学染色方法检测肝细胞癌组织芯片中Hsp90AB1的表达。运用生物信息学技术分析肝细胞癌相关基因芯片GSE54238;在数据库GEPIA中分析Hsp90AB1基因在正常肝组织及肝癌组织中的表达,对肝癌患者根据Hsp90AB1的表达差异进行生存分析。qRT-PCR检测Hsp90AB1基因在正常肝细胞系及肝癌细胞系中的表达。结果:免疫组化实验发现,与正常肝组织相比,Hsp90AB1在肝癌组织中表达上调(P<0.05)。对基因芯片GSE54238的生物信息学分析显示Hsp90AB1基因在正常肝组织和肝癌组织中的表达存在差异。GEPIA数据库分析发现Hsp90AB1基因在肝癌组织中显著过表达(P<0.05)。生存分析显示,Hsp90AB1高表达的肝癌患者预后相对较差(P<0.01)。相对于正常肝细胞系,Hsp90AB1在肝癌细胞系中高表达(P<0.05)。结论:Hsp90AB1基因可能是肝细胞癌潜在的促癌基因,其高表达可能与肝细胞癌患者较差的预后相关。展开更多
Beishashen(BSS)and Maidong(MD)are commonly used Medicine right for the treatment of non-small cell lung cancer(NSCLC),but their specific mechanism of action is not clear.In this study,network pharmacology and molecula...Beishashen(BSS)and Maidong(MD)are commonly used Medicine right for the treatment of non-small cell lung cancer(NSCLC),but their specific mechanism of action is not clear.In this study,network pharmacology and molecular docking techniques were used to investigate the molecular mechanisms of the therapeutic effects of BSS-MD on NSCLC and to experimentally validate some of the targets.The network pharmacology approach,including active ingredient and target screening,drug-compound-target network construction,protein-protein interaction(PPI)network,enrichment analysis,and molecular docking,was used to investigate the mechanism of action of Beisashen and Maitong on NSCLC.First,the active components of BSS-MD and their targets were predicted,of which 423 targets interacted with NSCLC targets.Then,network pharmacology showed that Stigmasterol,Quercetin,Alloisoimperatorin,Isoimperatorin,Beta-sitosterol were the core components of BSS-MD,and PLK1,HSP90AB1,and CDK1 were the key therapeutic targets.KEGG enrichment analysis indicated that the mechanism of action of BSS-MD in NSCLC treatment was related to the cell cycle.Then we further performed experimental validation.CCK-8 assay showed that BSS-MD inhibited LEWIS cell viability and promoted apoptosis in a dose-dependent manner.qPCR assay,immunofluorescence,and protein blotting experiments demonstrated that compared with the control group and the control group,the expression of PLK1,HSP90AB1,and CDK1 mRNAs and proteins were reduced in the treatment group(P<0.01).Therefore,we conclude that BSS-MD can block cell cycle progression by inhibiting the expression of PLK1,CDK1,and HSP90AB1 mRNAs and proteins to inhibit lung cancer cell growth and promote apoptosis,and emphasize that BSS-MD are promising adjuvants for NSCLC treatment.展开更多
文摘背景与目的热休克蛋白90AB1(heat shock protein 90 k Da alpha,class B member 1,Hsp90AB1)是ATP依赖的高度保守的分子伴侣,在多种肿瘤细胞中过表达。在肿瘤发生发展的信号传导通路中起着重要作用的一些分子,如表皮生长因子受体(epidermal growth factor receptor,EGFR)、人类表皮生长因子受体-2(human epidermal growth factor receptor-2,HER2)等,均是Hsp90AB1的底物蛋白。Hsp90AB1与这些底物蛋白相互作用并参与细胞的多种病理生理过程。本研究通过检测Hsp90AB1在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中的蛋白表达情况以初步探讨其临床意义。方法采用组织微阵列和免疫组织化学染色的方法检测Hsp90AB1在213例NSCLC及相应癌旁正常肺组织中的蛋白表达,并分析Hsp90AB1的表达与NSCLC临床病理参数及患者预后的关系。结果 Hsp90AB1在肺癌组织中的表达水平(阳性率54.0%)高于在正常肺组织中的表达水平(阳性率0.0%,P<0.001)。Hsp90AB1在肺腺癌中的表达阳性率为61.2%,高于肺鳞癌组织37.9%(P=0.002),并且其高表达与肺腺癌患者的不良预后相关(P=0.032)。Hsp90AB1蛋白表达水平与临床分期、淋巴结转移、病理分级等因素无关(P>0.05)。结论 Hsp90AB1在NSCLC组织中高表达,并且其表达水平与肺癌的病理类型及肺腺癌患者总生存期相关。
基金supported by the Pearl River S&T Nova Program of Guangzhou(201806010047)National Natural Science Foundation of China(32173001,3210284,31771587)+1 种基金China Postdoctoral Science Foundation Funded Project(2021M693678)Natural Science Foundation of Guangxi Province(2021GXNSFDA075015)。
文摘As a highly important fish virus,nervous necrosis virus(NNV)has caused severe economic losses to the aquaculture industry worldwide.Autophagy,an evolutionarily conserved intracellular degradation process,is involved in the pathogenesis of several viruses.Although NNV can induce autophagy to facilitate infection in grouper fish spleen cells,how it initiates and mediates autophagy pathways during the initial stage of infection is still unclear.Here,we found that red-spotted grouper NNV(RGNNV)induced autophagosome formation in two fish cell lines at 1.5 and 3 h post infection,indicating that autophagy is activated upon entry of RGNNV.Moreover,autophagic detection showed that RGNNV entry induced incomplete autophagy by impairing the fusion of autophagosomes with lysosomes.Further investigation revealed that binding of the RGNNV capsid protein(CP)to the Lateolabrax japonicus heat shock protein HSP90ab1(LjHSP90ab1),a cell surface receptor of RGNNV,contributed to RGNNV invasion-induced autophagy.Finally,we found that CP blocked the interaction of L.japonicus protein kinase B(AKT)with LjHSP90ab1 by competitively binding the NM domain of LjHSP90ab1 to inhibit the AKT-mechanistic target of the rapamycin(MTOR)pathway.This study provides novel insight into the relationship between NNV receptors and autophagy,which may help clarify the pathogenesis of NNV.
文摘目的:探讨热休克蛋白90ABl(heat shock protein 90 kDa alpha,class B member 1,Hsp90AB1)在肝细胞癌中的表达意义。方法:使用免疫组织化学染色方法检测肝细胞癌组织芯片中Hsp90AB1的表达。运用生物信息学技术分析肝细胞癌相关基因芯片GSE54238;在数据库GEPIA中分析Hsp90AB1基因在正常肝组织及肝癌组织中的表达,对肝癌患者根据Hsp90AB1的表达差异进行生存分析。qRT-PCR检测Hsp90AB1基因在正常肝细胞系及肝癌细胞系中的表达。结果:免疫组化实验发现,与正常肝组织相比,Hsp90AB1在肝癌组织中表达上调(P<0.05)。对基因芯片GSE54238的生物信息学分析显示Hsp90AB1基因在正常肝组织和肝癌组织中的表达存在差异。GEPIA数据库分析发现Hsp90AB1基因在肝癌组织中显著过表达(P<0.05)。生存分析显示,Hsp90AB1高表达的肝癌患者预后相对较差(P<0.01)。相对于正常肝细胞系,Hsp90AB1在肝癌细胞系中高表达(P<0.05)。结论:Hsp90AB1基因可能是肝细胞癌潜在的促癌基因,其高表达可能与肝细胞癌患者较差的预后相关。
基金This research was supported by“Zhejiang Province Chinese Medicine Science and Technology Program Key Projects”(No.2021ZZ008).
文摘Beishashen(BSS)and Maidong(MD)are commonly used Medicine right for the treatment of non-small cell lung cancer(NSCLC),but their specific mechanism of action is not clear.In this study,network pharmacology and molecular docking techniques were used to investigate the molecular mechanisms of the therapeutic effects of BSS-MD on NSCLC and to experimentally validate some of the targets.The network pharmacology approach,including active ingredient and target screening,drug-compound-target network construction,protein-protein interaction(PPI)network,enrichment analysis,and molecular docking,was used to investigate the mechanism of action of Beisashen and Maitong on NSCLC.First,the active components of BSS-MD and their targets were predicted,of which 423 targets interacted with NSCLC targets.Then,network pharmacology showed that Stigmasterol,Quercetin,Alloisoimperatorin,Isoimperatorin,Beta-sitosterol were the core components of BSS-MD,and PLK1,HSP90AB1,and CDK1 were the key therapeutic targets.KEGG enrichment analysis indicated that the mechanism of action of BSS-MD in NSCLC treatment was related to the cell cycle.Then we further performed experimental validation.CCK-8 assay showed that BSS-MD inhibited LEWIS cell viability and promoted apoptosis in a dose-dependent manner.qPCR assay,immunofluorescence,and protein blotting experiments demonstrated that compared with the control group and the control group,the expression of PLK1,HSP90AB1,and CDK1 mRNAs and proteins were reduced in the treatment group(P<0.01).Therefore,we conclude that BSS-MD can block cell cycle progression by inhibiting the expression of PLK1,CDK1,and HSP90AB1 mRNAs and proteins to inhibit lung cancer cell growth and promote apoptosis,and emphasize that BSS-MD are promising adjuvants for NSCLC treatment.
