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
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
目的:通过高通量转录组测序和相对定量血清蛋白组学技术筛选肝癌转移相关的关键基因.方法:利用Ion Proton.测序仪比较人肝癌细胞株(Smmc-7721)和正常人肝细胞株(L-02)的差异表达基因,对差异表达基因进行聚类分析、GO功能注释和富集分析...目的:通过高通量转录组测序和相对定量血清蛋白组学技术筛选肝癌转移相关的关键基因.方法:利用Ion Proton.测序仪比较人肝癌细胞株(Smmc-7721)和正常人肝细胞株(L-02)的差异表达基因,对差异表达基因进行聚类分析、GO功能注释和富集分析,获得肝癌细胞转移相关基因;收集10例肝细胞癌(hepatocellular carcinoma,HCC)患者血清及匹配10例正常人血清,通过相对和绝对定量的同位素标记(isobaric tags for relative and absolute quantitation,iTRAQ)联合基质辅助激光解吸电离串联飞行时间质谱(ma t r ixassisted laser desorption/ionization tandemtime of flight mass spectrometry,MALDITOF/MS)检测肝癌与正常对照组血清差异表达蛋白;将两组学结果进行交集分析,确定肝癌转移关键基因,并在76例HCC和癌旁组织中进行免疫组织化学验证.结果:对肝癌细胞及正常肝细胞进行转录组测序分析,共得到肝癌细胞差异表达基因618个,生物信息学分析差异表达基因主要聚集在转移相关、转录因子相关、氧化还原过程等14个分子功能,其中转移相关功能基因所占比例最大,达15.05%(93/618);血清蛋白组学分析共得到69个肝癌血清差异表达蛋白,其中在肝癌组上调33个,下调36个;将转录组测序筛选的与转移功能相关基因与蛋白质组学进行交集分析,发现有3个共同交集的差异因子,其中差异最明显的是肝癌中表达上调的热休克蛋白90 AA1(heat shock protein 90 AA1,HSP90AA1);免疫组织化学验证结果显示,HSP90α表达强阳性在门脉转移和无门脉转移HCC组织中的比例分别为66.7%(16/24)和25%(13/52)(P<0.005),HSP90α在有门脉转移的肝癌组织表达明显增高.结论:利用转录组结合血清蛋白组策略,发现HSP90AA1可能是在肝癌转移重要基因.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.
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.
文摘目的:通过高通量转录组测序和相对定量血清蛋白组学技术筛选肝癌转移相关的关键基因.方法:利用Ion Proton.测序仪比较人肝癌细胞株(Smmc-7721)和正常人肝细胞株(L-02)的差异表达基因,对差异表达基因进行聚类分析、GO功能注释和富集分析,获得肝癌细胞转移相关基因;收集10例肝细胞癌(hepatocellular carcinoma,HCC)患者血清及匹配10例正常人血清,通过相对和绝对定量的同位素标记(isobaric tags for relative and absolute quantitation,iTRAQ)联合基质辅助激光解吸电离串联飞行时间质谱(ma t r ixassisted laser desorption/ionization tandemtime of flight mass spectrometry,MALDITOF/MS)检测肝癌与正常对照组血清差异表达蛋白;将两组学结果进行交集分析,确定肝癌转移关键基因,并在76例HCC和癌旁组织中进行免疫组织化学验证.结果:对肝癌细胞及正常肝细胞进行转录组测序分析,共得到肝癌细胞差异表达基因618个,生物信息学分析差异表达基因主要聚集在转移相关、转录因子相关、氧化还原过程等14个分子功能,其中转移相关功能基因所占比例最大,达15.05%(93/618);血清蛋白组学分析共得到69个肝癌血清差异表达蛋白,其中在肝癌组上调33个,下调36个;将转录组测序筛选的与转移功能相关基因与蛋白质组学进行交集分析,发现有3个共同交集的差异因子,其中差异最明显的是肝癌中表达上调的热休克蛋白90 AA1(heat shock protein 90 AA1,HSP90AA1);免疫组织化学验证结果显示,HSP90α表达强阳性在门脉转移和无门脉转移HCC组织中的比例分别为66.7%(16/24)和25%(13/52)(P<0.005),HSP90α在有门脉转移的肝癌组织表达明显增高.结论:利用转录组结合血清蛋白组策略,发现HSP90AA1可能是在肝癌转移重要基因.
