Lymphatic metastasis(LM)emerges as an independent prognostic marker for hypopharyngeal squamous cell carcinoma(HSPSCC),chiefly contributing to treatment inefficacy.This study aimed to scrutinize the prognostic relevan...Lymphatic metastasis(LM)emerges as an independent prognostic marker for hypopharyngeal squamous cell carcinoma(HSPSCC),chiefly contributing to treatment inefficacy.This study aimed to scrutinize the prognostic relevance of HSP90AA1 and its potential regulatory mechanism of concerning LM in HPSCC.Methods:In a preceding investigation,HSP90AA1,a differential gene,was discovered through transcriptome sequencing of HPSCC tissues,considering both the presence and absence of LM.Validation of HSP90AA1 expression was accomplished via qRT-PCR,western-blotting(WB),and immunohistochemistry(IHC),while its prognostic significance was assessed employing Kaplan–Meier survival analysis(KMSA),log-rank test(LR),and Cox’s regression analysis(CRA).Bioinformatics techniques facilitated the prediction and analysis of its plausible mechanisms in LM,further substantiated by in vitro and in vivo experiments utilizing FaDu cell lines.Results:HSP90AA1 is substantially upregulated in HPSCC with LM and is identified as an independent prognostic risk determinant.The down-regulation of HSP90AA1 can achieve inhibition of tumor cell proliferation,migration and invasion.Both in vivo experiments and Bioinformatics exploration hint at promoting LM by Epithelial-mesenchymal transition(EMT),regulated by HSP90AA1.Conclusions:HSP90AA1,by controlling EMT,can foster LM in HPSCC.This finding sets the foundation for delving into new therapeutic targets for HPSCC.展开更多
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
HSP90 AA1 is part of the heat shock protein 90 gene family and has important functions against heat stress. We report a case of molecular level parallel evolution of the HSP90 AA1 gene in high elevation amphibians. HS...HSP90 AA1 is part of the heat shock protein 90 gene family and has important functions against heat stress. We report a case of molecular level parallel evolution of the HSP90 AA1 gene in high elevation amphibians. HSP90 AA1 gene sequences of four high-elevation anurans, Bufo gargarizans, Nanorana parkeri, Rana kukunoris, and Scutiger boulengeri, were compared along with five of their low-elevation relatives. A total of 16 amino-acid sites were identified as parallel evolution between N. parkeri and R. kukunoris. We generated both model based（Zhang and Kumar＇s test） and empirical data based（parallel/divergence plotting） null distributions for non-parallel evolution, and both methods clearly determined that the observed number of parallel substitutions were significantly more than the null expectation. Furthermore, on the HSP90 AA1 gene tree, N. parkeri and R. kukunoris formed a strongly supported clade that was away from their respective relatives. This study provides a clear case of molecular parallel evolution, which may have significant implications in understanding the genetic mechanisms of high-elevation adaptation.展开更多
Background： The purpose of this study was to determine the association of three proteins involved in sperm function on the freezability of porcine semen： the heat shock protein 90 alpha（HSP90a）, the Niemann-Pick d...Background： The purpose of this study was to determine the association of three proteins involved in sperm function on the freezability of porcine semen： the heat shock protein 90 alpha（HSP90a）, the Niemann-Pick disease type C2 protein（NPC2）, and lipocalin-type prostaglandin D synthase（L-PGDS）.Six adult boars（each boar was ejaculated three times, 18 in total） were classified by freezability based on the percentage of functionally competent sperm. The male semen with highest freezability（MHF） and the male semen with lowest freezability（MLF） were centrifuged immediately after collection to separate seminal plasma and spermatozoa to make four possible combinations of these two components and to incubate them for 3 h,adjusting the temperature to 17 °C, to freeze them afterwards. The quantification of proteins was performed in two stages： at zero and at 3 h after incubation of the four combinations.Results： The spermatozoa × incubation time（IT） interaction only had effect（P 〈 0.01） on HSP90 a levels; this protein increased in seminal plasma, after 3 h of incubation, in larger quantity（P 〈 0.05） in combinations with MLF spermatozoa. In relation with the NPC2 protein, two isoforms of 16 and 19 kDa were identified. The 19 kDa isoform was affected（P 〈 0.01） only by the seminal plasma × IT interaction, with superior values（P 〈 0.01） both at zero and three hours of incubation, in the combinations with MHF seminal plasma; and 16 kDa isoform was affected（P 〈 0.01） only by the IT with reduction after 3 h of incubation. The levels of L-PGDS was affected（P 〈 0.01）only by the spermatozoa × IT interaction, which reduced（P 〈 0.01） in combinations with MLF spermatozoa after 3 h of incubation.Conclusions： It is possible to consider that the three proteins evaluated were associated with freezability of boar semen due, especially, to the fact that mixtures with MLF spermatozoa showed greater increase levels of the HSP90 a protein and reduction of L-PGDS in plasma. In addition, the seminal plasma of MHF had higher concentration of the NPC2 of 19 kDa protein, which was reduced by incubating with MHF spermatozoa.展开更多
Methamphetamine(METH)is one of the most widely abused synthetic drugs in the world,with the abuser present hyperthermia(HT)and psychiatric symptoms.However,the mechanism involved in METH/HT-induced neurotoxicity remai...Methamphetamine(METH)is one of the most widely abused synthetic drugs in the world,with the abuser present hyperthermia(HT)and psychiatric symptoms.However,the mechanism involved in METH/HT-induced neurotoxicity remains elusive.Here,we investigated the role of heat shock protein 90 alpha(HSP90a)in METH/HT(39.5C)-induced necroptosis in rat striatum neurons and rat in vivo model.Western blot showed that the expression of HSP90a increased,and HSP90a specific inhibitor geldanamycin(GA)and shRNA of HSP90a could attenuate METH/HT-induced upregulation of receptor interacting protein 3(RIP3),pRIP3,mixed lineage kinase domain-like protein(MLKL)and pMLKL in cultured striatum neurons and in vivo model.Propidium iodide staining and lactate dehydrogenase assay results showed that METH/HT-induced necroptosis is partially inhibited by both necrostain-1,GA and shRNA of HSP90a in vitro and in vivo model.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.82173303)Natural Science Foundation of Chongqing,China(Grant No.cstc2021ycjh-bgzxm0149).
文摘Lymphatic metastasis(LM)emerges as an independent prognostic marker for hypopharyngeal squamous cell carcinoma(HSPSCC),chiefly contributing to treatment inefficacy.This study aimed to scrutinize the prognostic relevance of HSP90AA1 and its potential regulatory mechanism of concerning LM in HPSCC.Methods:In a preceding investigation,HSP90AA1,a differential gene,was discovered through transcriptome sequencing of HPSCC tissues,considering both the presence and absence of LM.Validation of HSP90AA1 expression was accomplished via qRT-PCR,western-blotting(WB),and immunohistochemistry(IHC),while its prognostic significance was assessed employing Kaplan–Meier survival analysis(KMSA),log-rank test(LR),and Cox’s regression analysis(CRA).Bioinformatics techniques facilitated the prediction and analysis of its plausible mechanisms in LM,further substantiated by in vitro and in vivo experiments utilizing FaDu cell lines.Results:HSP90AA1 is substantially upregulated in HPSCC with LM and is identified as an independent prognostic risk determinant.The down-regulation of HSP90AA1 can achieve inhibition of tumor cell proliferation,migration and invasion.Both in vivo experiments and Bioinformatics exploration hint at promoting LM by Epithelial-mesenchymal transition(EMT),regulated by HSP90AA1.Conclusions:HSP90AA1,by controlling EMT,can foster LM in HPSCC.This finding sets the foundation for delving into new therapeutic targets for HPSCC.
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.
基金supported by the National Nature Science Foundation of China (grant number 31328021 to Jinzhong FU)NSERC of Canada (a discovery grant to Jinzhong FU)
文摘HSP90 AA1 is part of the heat shock protein 90 gene family and has important functions against heat stress. We report a case of molecular level parallel evolution of the HSP90 AA1 gene in high elevation amphibians. HSP90 AA1 gene sequences of four high-elevation anurans, Bufo gargarizans, Nanorana parkeri, Rana kukunoris, and Scutiger boulengeri, were compared along with five of their low-elevation relatives. A total of 16 amino-acid sites were identified as parallel evolution between N. parkeri and R. kukunoris. We generated both model based（Zhang and Kumar＇s test） and empirical data based（parallel/divergence plotting） null distributions for non-parallel evolution, and both methods clearly determined that the observed number of parallel substitutions were significantly more than the null expectation. Furthermore, on the HSP90 AA1 gene tree, N. parkeri and R. kukunoris formed a strongly supported clade that was away from their respective relatives. This study provides a clear case of molecular parallel evolution, which may have significant implications in understanding the genetic mechanisms of high-elevation adaptation.
基金funded by the Vice-Rector’s Office at Universidad de Caldas
文摘Background： The purpose of this study was to determine the association of three proteins involved in sperm function on the freezability of porcine semen： the heat shock protein 90 alpha（HSP90a）, the Niemann-Pick disease type C2 protein（NPC2）, and lipocalin-type prostaglandin D synthase（L-PGDS）.Six adult boars（each boar was ejaculated three times, 18 in total） were classified by freezability based on the percentage of functionally competent sperm. The male semen with highest freezability（MHF） and the male semen with lowest freezability（MLF） were centrifuged immediately after collection to separate seminal plasma and spermatozoa to make four possible combinations of these two components and to incubate them for 3 h,adjusting the temperature to 17 °C, to freeze them afterwards. The quantification of proteins was performed in two stages： at zero and at 3 h after incubation of the four combinations.Results： The spermatozoa × incubation time（IT） interaction only had effect（P 〈 0.01） on HSP90 a levels; this protein increased in seminal plasma, after 3 h of incubation, in larger quantity（P 〈 0.05） in combinations with MLF spermatozoa. In relation with the NPC2 protein, two isoforms of 16 and 19 kDa were identified. The 19 kDa isoform was affected（P 〈 0.01） only by the seminal plasma × IT interaction, with superior values（P 〈 0.01） both at zero and three hours of incubation, in the combinations with MHF seminal plasma; and 16 kDa isoform was affected（P 〈 0.01） only by the IT with reduction after 3 h of incubation. The levels of L-PGDS was affected（P 〈 0.01）only by the spermatozoa × IT interaction, which reduced（P 〈 0.01） in combinations with MLF spermatozoa after 3 h of incubation.Conclusions： It is possible to consider that the three proteins evaluated were associated with freezability of boar semen due, especially, to the fact that mixtures with MLF spermatozoa showed greater increase levels of the HSP90 a protein and reduction of L-PGDS in plasma. In addition, the seminal plasma of MHF had higher concentration of the NPC2 of 19 kDa protein, which was reduced by incubating with MHF spermatozoa.
文摘Methamphetamine(METH)is one of the most widely abused synthetic drugs in the world,with the abuser present hyperthermia(HT)and psychiatric symptoms.However,the mechanism involved in METH/HT-induced neurotoxicity remains elusive.Here,we investigated the role of heat shock protein 90 alpha(HSP90a)in METH/HT(39.5C)-induced necroptosis in rat striatum neurons and rat in vivo model.Western blot showed that the expression of HSP90a increased,and HSP90a specific inhibitor geldanamycin(GA)and shRNA of HSP90a could attenuate METH/HT-induced upregulation of receptor interacting protein 3(RIP3),pRIP3,mixed lineage kinase domain-like protein(MLKL)and pMLKL in cultured striatum neurons and in vivo model.Propidium iodide staining and lactate dehydrogenase assay results showed that METH/HT-induced necroptosis is partially inhibited by both necrostain-1,GA and shRNA of HSP90a in vitro and in vivo model.
