It has been shown that the high expression of human epididymis protein 4(HE4)in most lung cancers is related to the poor prognosis of patients,but the mechanism of pathological transformation of HE4 in lung cancer is ...It has been shown that the high expression of human epididymis protein 4(HE4)in most lung cancers is related to the poor prognosis of patients,but the mechanism of pathological transformation of HE4 in lung cancer is still unclear.The current study is expected to clarify the function and mechanism of HE4 in the occurrence and metastasis of lung adenocarcinoma(LUAD).Immunoblotting evaluated HE4 expression in lung cancer cell lines and biopsies,and through analysis of The Cancer Genome Atlas(TCGA)dataset.Frequent HE4 overexpression was demonstrated in LUAD,but not in lung squamous cell carcinoma(LUSC),indicating that HE4 can serve as a biomarker to distinguish between LUAD and LUSC.HE4 knockdown significantly inhibited cell growth,colony formation,wound healing,and invasion,and blocked the G1-phase of the cell cycle in LUAD cell lines through inactivation of the EGFR signaling downstream including PI3K/AKT/mTOR and RAF/MAPK pathways.The first-line EGFR inhibitor gefitinib and HE4 shRNA had no synergistic inhibitory effect on the growth of lung adenocarcinoma cells,while the third-line EGFR inhibitor osimertinib showed additive anti-proliferative effects.Moreover,we provided evidence that HE4 regulated EGFR expression by transcription regulation and protein interaction in LUAD.Our findings suggest that HE4 positively modulates the EGFR signaling pathway to promote growth and invasiveness in LUAD and highlight that targeting HE4 could be a novel strategy for LUAD treatment.展开更多
BACKGROUND Heat shock protein A4(HSPA4)belongs to molecular chaperone protein family which plays important roles within variable cellular activities,including cancer initiation and progression.However,the prognostic a...BACKGROUND Heat shock protein A4(HSPA4)belongs to molecular chaperone protein family which plays important roles within variable cellular activities,including cancer initiation and progression.However,the prognostic and immunological significance of HSPA4 in lung adenocarcinoma(LUAD)has not been revealed yet.AIM To explore the prognostic and immunological roles of HSPA4 to identify a novel prognostic biomarker and therapeutic target for LUAD.METHODS We assessed the prognostic and immunological significance of HSPA4 in LUAD using data from The Cancer Genome Atlas database.The association between HSPA4 expression and clinical-pathological features was assessed through Kruskal-Wallis and Wilcoxon signed-rank test.Univariate/multivariate Cox regression analyses and Kaplan-Meier curves were employed to evaluate prognostic factors,including HSPA4,in LUAD.Gene set enrichment analysis(GSEA)was conducted to identify the key signaling pathways associated with HSPA4.The correlation between HSPA4 expression and cancer immune infiltration was evaluated using single-sample gene set enrichment analysis(ssGSEA).RESULTS Overexpressing HSPA4 was significantly related to advanced pathologic TNM stage,advanced pathologic stage,progression disease status of primary therapy outcome and female subgroups with LUAD.In addition,increased HSPA4 expression was found to be related to worse disease-specific survival and overall survival.GSEA analysis indicated a significant correlation between HSPA4 and cell cycle regulation and immune response,particularly through diminishing the function of cytotoxicity cells and CD8 T cells.The ssGSEA algorithm showed a positive correlation between HSPA4 expression and infiltrating levels of Th2 cells,while a negative correlation was observed with cytotoxic cell infiltration levels.CONCLUSION Our findings indicate HSPA4 is related to prognosis and immune cell infiltrates and may act as a novel prognostic biomarker and therapeutic target for LUAD.展开更多
目的:微小染色体维持蛋白4(minichromosome maintenance protein 4,MCM4)是DNA复制起始和延伸阶段的调控因子。本研究旨在探究肺腺癌(lung adenocarcinoma,LUAD)中MCM4的表达与肿瘤微环境(tumor microenvironment,TME)中免疫细胞和分子...目的:微小染色体维持蛋白4(minichromosome maintenance protein 4,MCM4)是DNA复制起始和延伸阶段的调控因子。本研究旨在探究肺腺癌(lung adenocarcinoma,LUAD)中MCM4的表达与肿瘤微环境(tumor microenvironment,TME)中免疫细胞和分子的相关性,以及MCM4预测患者免疫治疗敏感性和预后的能力。方法:从癌症基因组图谱(The Cancer Genome Atlas,TCGA)、基因表达综合(Gene Expression Omnibus,GEO)数据库中下载并筛选LUAD患者的转录组数据和临床数据进行分析。其中TCGA数据集的503例患者为训练集,GEO数据集的857例患者为验证集。采用TIMER(Tumor Immune Estimation Resource)数据库对MCM4在正常组织和肿瘤组织中的表达情况进行分析。将LUAD患者肿瘤样本划分为MCM4低表达组和MCM4高表达组,采用R软件分析LUAD患者MCM4表达与总生存期(overall survival,OS)的关系;采用京都基因与基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)中的基因集进行基因富集分析,筛选与MCM4基因表达相关的生物学过程;采用肿瘤免疫功能障碍与排斥(tumor immune dysfunction and exclusion,TIDE)评分预测患者的免疫治疗的效果;评估LUAD患者TME的免疫评分、基质评分和ESTIMATE(estimation of stromal and immune cells in malignant tumor tissues using expression data)评分,分析MCM4表达与TME中免疫细胞、免疫检查点(immune checkpoint,ICs)之间的关系。收集武汉大学人民医院病理确诊为LUAD的穿刺或者手术切片60张(55张为LUAD的癌组织和癌旁组织,5张为正常肺组织)及临床病理资料,采用免疫组织化学染色法进行临床验证。结果:MCM4基因和蛋白质在肺正常组织和腺癌组织中差异表达。在训练集和验证集中,MCM4低表达组患者的OS均优于MCM4高表达组;MCM4均可以影响LUAD患者的预后;MCM4表达与卵母细胞减数分裂、细胞周期和DNA复制通路显著相关,且MCM4与Ki-67编码基因MKI67和增殖相关基因C1orf112均呈显著正相关;MCM4表达与TIDE评分均呈负相关,与ICs表达呈正相关,说明MCM4的表达可预测患者的免疫治疗效果;与MCM4低表达组比较,MCM4高表达组的肿瘤纯度均较高,基质细胞和免疫细胞比例均较低,且MCM4表达与CD8+T细胞均呈负相关,与Th2、骨髓来源抑制性细胞均呈显著正相关,说明肿瘤组织的MCM4高表达与患者预后不佳相关。泛癌分析结果表明:在33种肿瘤中,大部分肿瘤的MCM4表达与ICs、Th2细胞、调节性T细胞、程序性死亡受体1(programmed death-1,PD-1)、程序性死亡受体配体1(programmed death-ligand 1,PD-L1)的表达及PD-1-PD-L1评分均呈正相关,与CD8^(+)T细胞、NK细胞均呈负相关。临床数据验证结果表明:MCM4在肿瘤组织中的染色强度和阳性细胞占比高于肿瘤间质,肿瘤组织的MCM4表达水平高于正常组织;MCM4评分与Ki-67呈显著正相关。结论:MCM4作为增殖信号影响肺腺癌的免疫浸润和预后,且MCM4高表达与TME内抑制性免疫细胞浸润及免疫治疗获益概率升高相关。展开更多
基金supported by National Natural Science Foundation of China(82272695)the Key Program of Natural Science Foundation of Zhejiang Province(LZ23H160004)National Undergraduate Training Program for Innovation,and Zhejiang Xinmiao Talents Program,China.
文摘It has been shown that the high expression of human epididymis protein 4(HE4)in most lung cancers is related to the poor prognosis of patients,but the mechanism of pathological transformation of HE4 in lung cancer is still unclear.The current study is expected to clarify the function and mechanism of HE4 in the occurrence and metastasis of lung adenocarcinoma(LUAD).Immunoblotting evaluated HE4 expression in lung cancer cell lines and biopsies,and through analysis of The Cancer Genome Atlas(TCGA)dataset.Frequent HE4 overexpression was demonstrated in LUAD,but not in lung squamous cell carcinoma(LUSC),indicating that HE4 can serve as a biomarker to distinguish between LUAD and LUSC.HE4 knockdown significantly inhibited cell growth,colony formation,wound healing,and invasion,and blocked the G1-phase of the cell cycle in LUAD cell lines through inactivation of the EGFR signaling downstream including PI3K/AKT/mTOR and RAF/MAPK pathways.The first-line EGFR inhibitor gefitinib and HE4 shRNA had no synergistic inhibitory effect on the growth of lung adenocarcinoma cells,while the third-line EGFR inhibitor osimertinib showed additive anti-proliferative effects.Moreover,we provided evidence that HE4 regulated EGFR expression by transcription regulation and protein interaction in LUAD.Our findings suggest that HE4 positively modulates the EGFR signaling pathway to promote growth and invasiveness in LUAD and highlight that targeting HE4 could be a novel strategy for LUAD treatment.
文摘BACKGROUND Heat shock protein A4(HSPA4)belongs to molecular chaperone protein family which plays important roles within variable cellular activities,including cancer initiation and progression.However,the prognostic and immunological significance of HSPA4 in lung adenocarcinoma(LUAD)has not been revealed yet.AIM To explore the prognostic and immunological roles of HSPA4 to identify a novel prognostic biomarker and therapeutic target for LUAD.METHODS We assessed the prognostic and immunological significance of HSPA4 in LUAD using data from The Cancer Genome Atlas database.The association between HSPA4 expression and clinical-pathological features was assessed through Kruskal-Wallis and Wilcoxon signed-rank test.Univariate/multivariate Cox regression analyses and Kaplan-Meier curves were employed to evaluate prognostic factors,including HSPA4,in LUAD.Gene set enrichment analysis(GSEA)was conducted to identify the key signaling pathways associated with HSPA4.The correlation between HSPA4 expression and cancer immune infiltration was evaluated using single-sample gene set enrichment analysis(ssGSEA).RESULTS Overexpressing HSPA4 was significantly related to advanced pathologic TNM stage,advanced pathologic stage,progression disease status of primary therapy outcome and female subgroups with LUAD.In addition,increased HSPA4 expression was found to be related to worse disease-specific survival and overall survival.GSEA analysis indicated a significant correlation between HSPA4 and cell cycle regulation and immune response,particularly through diminishing the function of cytotoxicity cells and CD8 T cells.The ssGSEA algorithm showed a positive correlation between HSPA4 expression and infiltrating levels of Th2 cells,while a negative correlation was observed with cytotoxic cell infiltration levels.CONCLUSION Our findings indicate HSPA4 is related to prognosis and immune cell infiltrates and may act as a novel prognostic biomarker and therapeutic target for LUAD.
文摘目的:微小染色体维持蛋白4(minichromosome maintenance protein 4,MCM4)是DNA复制起始和延伸阶段的调控因子。本研究旨在探究肺腺癌(lung adenocarcinoma,LUAD)中MCM4的表达与肿瘤微环境(tumor microenvironment,TME)中免疫细胞和分子的相关性,以及MCM4预测患者免疫治疗敏感性和预后的能力。方法:从癌症基因组图谱(The Cancer Genome Atlas,TCGA)、基因表达综合(Gene Expression Omnibus,GEO)数据库中下载并筛选LUAD患者的转录组数据和临床数据进行分析。其中TCGA数据集的503例患者为训练集,GEO数据集的857例患者为验证集。采用TIMER(Tumor Immune Estimation Resource)数据库对MCM4在正常组织和肿瘤组织中的表达情况进行分析。将LUAD患者肿瘤样本划分为MCM4低表达组和MCM4高表达组,采用R软件分析LUAD患者MCM4表达与总生存期(overall survival,OS)的关系;采用京都基因与基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)中的基因集进行基因富集分析,筛选与MCM4基因表达相关的生物学过程;采用肿瘤免疫功能障碍与排斥(tumor immune dysfunction and exclusion,TIDE)评分预测患者的免疫治疗的效果;评估LUAD患者TME的免疫评分、基质评分和ESTIMATE(estimation of stromal and immune cells in malignant tumor tissues using expression data)评分,分析MCM4表达与TME中免疫细胞、免疫检查点(immune checkpoint,ICs)之间的关系。收集武汉大学人民医院病理确诊为LUAD的穿刺或者手术切片60张(55张为LUAD的癌组织和癌旁组织,5张为正常肺组织)及临床病理资料,采用免疫组织化学染色法进行临床验证。结果:MCM4基因和蛋白质在肺正常组织和腺癌组织中差异表达。在训练集和验证集中,MCM4低表达组患者的OS均优于MCM4高表达组;MCM4均可以影响LUAD患者的预后;MCM4表达与卵母细胞减数分裂、细胞周期和DNA复制通路显著相关,且MCM4与Ki-67编码基因MKI67和增殖相关基因C1orf112均呈显著正相关;MCM4表达与TIDE评分均呈负相关,与ICs表达呈正相关,说明MCM4的表达可预测患者的免疫治疗效果;与MCM4低表达组比较,MCM4高表达组的肿瘤纯度均较高,基质细胞和免疫细胞比例均较低,且MCM4表达与CD8+T细胞均呈负相关,与Th2、骨髓来源抑制性细胞均呈显著正相关,说明肿瘤组织的MCM4高表达与患者预后不佳相关。泛癌分析结果表明:在33种肿瘤中,大部分肿瘤的MCM4表达与ICs、Th2细胞、调节性T细胞、程序性死亡受体1(programmed death-1,PD-1)、程序性死亡受体配体1(programmed death-ligand 1,PD-L1)的表达及PD-1-PD-L1评分均呈正相关,与CD8^(+)T细胞、NK细胞均呈负相关。临床数据验证结果表明:MCM4在肿瘤组织中的染色强度和阳性细胞占比高于肿瘤间质,肿瘤组织的MCM4表达水平高于正常组织;MCM4评分与Ki-67呈显著正相关。结论:MCM4作为增殖信号影响肺腺癌的免疫浸润和预后,且MCM4高表达与TME内抑制性免疫细胞浸润及免疫治疗获益概率升高相关。
