Objective:This study aimed at exploring the effects of the epigenetic regulator,chidamide,on reprogramming the immunosuppressive tumor microenvironment in small cell lung cancer(SCLC),particularly the roles in macroph...Objective:This study aimed at exploring the effects of the epigenetic regulator,chidamide,on reprogramming the immunosuppressive tumor microenvironment in small cell lung cancer(SCLC),particularly the roles in macrophage polarization and angiogenesis.The therapeutic efficacy of combining chidamide with the anti-angiogenic agent,anlotinib,for refractory SCLC was also evaluated.Methods:RNA sequencing and functional validation were performed to assess chidamide’s effects on macrophages.Signal transducer and activator of transcription 4(STAT4)-mediated transcriptional activation of CCL2 was confirmed with ChIP-qPCR.The synergistic efficacy of chidamide in combination with anlotinib was tested in preclinical models.Results:Chidamide enhanced macrophage infiltration and induced macrophage polarization toward the anti-tumor M1 phenotype.Mechanistically,chidamide upregulated CCL2 via STAT4 transcriptional activation,thereby reshaping the tumor immune microenvironment(TIME).Combining chidamide with anlotinib synergistically suppressed tumor growth and remodeled the immunosuppressive TME in SCLC in vivo.Conclusions:Chidamide reshaped the SCLC TIME by activating STAT4/CCL2,thus driving M1 macrophage polarization and enhancing anti-tumor immunity.Our findings highlight coordinated TIME-targeted therapy as a translatable strategy to overcome therapeutic resistance in SCLC and provide a rationale for clinical trials examining epigenetic and anti-angiogenic therapeutics combinations.展开更多
目的观察喘可治注射液(CKZ)对慢性阻塞性肺疾病(COPD)大鼠肺组织IL-12/IL-12R/STAT4信号通路的影响。方法 50只SD大鼠随机分为正常组,模型组,CKZ高、中、低剂量(CKZ-H、CHZ-M、CKZ-L)组。除正常组外,各组复制COPD大鼠模型,并给予相应药...目的观察喘可治注射液(CKZ)对慢性阻塞性肺疾病(COPD)大鼠肺组织IL-12/IL-12R/STAT4信号通路的影响。方法 50只SD大鼠随机分为正常组,模型组,CKZ高、中、低剂量(CKZ-H、CHZ-M、CKZ-L)组。除正常组外,各组复制COPD大鼠模型,并给予相应药物治疗,观察CKZ对COPD大鼠血清和肺泡灌洗液(BALF)中γ-干扰素(IFN-γ)、白细胞介素12(IL-12)、白细胞介素12受体(IL-12R)以及肺组织信号转导子和转录激活子4(STAT4)m RNA表达的影响。结果喘可治注射液各剂量组肺组织病理形态均有不同程度的改善,且降低COPD大鼠血清和BALF中IFN-γ、IL-12、IL-12R(除CKZ-L组BALF中IL-12R外)水平(P<0.05,P<0.01),可抑制大鼠肺组织STAT4 m RNA的表达(P<0.01,P<0.05)。结论喘可治注射液能一定程度改善COPD大鼠肺组织病理损害,其作用机制可能与其抑制IFN-γ、IL-12、IL-12R表达,干扰IL-12/IL-12R/STAT4信号通路对靶基因的激活,下调STAT4 m RNA表达,进而调节Th1/Th2分化失衡有关。展开更多
为了阐明水牛信号转导子和转录激活子4(signal transducer and activator of transcription 4,STAT4)基因在水牛卵泡、胚胎发生及泌乳调节过程中的作用及分子机制,本试验采用了3′-RACE克隆获得STAT4基因,并对其核苷酸序列和蛋白质序列...为了阐明水牛信号转导子和转录激活子4(signal transducer and activator of transcription 4,STAT4)基因在水牛卵泡、胚胎发生及泌乳调节过程中的作用及分子机制,本试验采用了3′-RACE克隆获得STAT4基因,并对其核苷酸序列和蛋白质序列进行了信息学分析,通过构建其真核表达载体并转染HEK293T细胞验证所构建载体的准确性。结果表明,水牛STAT4基因编码区长2 247bp,3′-UTR区长268bp,编码748个氨基酸。BLAST分析显示,水牛STAT4核苷酸序列与牛、山羊、绵羊、猪、马、犬和人相应序列的同源性分别为99%、99%、99%、95%、93%、93%和92%,系统进化树分析结果表明,STAT4基因在不同物种及进化的过程中具有高度保守性。蛋白质分析结果表明,STAT4蛋白呈弱酸性,无信号肽,定位于细胞质,存在STAT_int、STAT_alpha、STAT_bind和SH2_STAT4等结构域。microRNA靶标预测显示bta-miR-200a、bta-miR-2429和bta-miR-2410等为STAT4 3′-UTR潜在microRNAs。试验成功构建了水牛STAT4基因真核表达载体pEGFPN1-STAT4,转染HEK293T后产生较强的绿色荧光信号,表明能够形成STAT4-EGFP融合蛋白。展开更多
目的:整理挖掘TCGA(the Cancer Genome Atlas)和GEO(gene expression omnibus)数据库中乳腺癌基因表达数据集的相关数据,分析STAT4基因在乳腺癌中的表达及临床意义,探讨丹参酮ⅡA对乳腺癌细胞中STAT4基因的干预效果。方法:下载TCGA数据...目的:整理挖掘TCGA(the Cancer Genome Atlas)和GEO(gene expression omnibus)数据库中乳腺癌基因表达数据集的相关数据,分析STAT4基因在乳腺癌中的表达及临床意义,探讨丹参酮ⅡA对乳腺癌细胞中STAT4基因的干预效果。方法:下载TCGA数据库中的乳腺癌数据集(TCGA Breast Cancer(BRCA))和GEO数据库中有关乳腺癌和丹参酮ⅡA干预乳腺癌的数据集(GSE42586、GSE85871),对所获取的数据资料进行整理并作统计分析,通过免疫印迹实验和分子对接对数据分析结果进行验证。结果:对TCGA和GEO数据库中所有关于乳腺癌和正常组织乳腺的STAT4基因表达数据进行统计分析,结果发现乳腺癌组织中STAT4基因的表达明显低于正常乳腺组织,差异有统计学意义(P<0.05)。通过生存分析发现高表达STAT4基因的乳腺癌患者无病生存期明显好于低表达者,高表达患者的预后更好(P<0.05)。GSE85871数据集中有关丹参酮ⅡA作用于乳腺癌MCF7细胞的基因表达数据表明丹参酮ⅡA可以上调MCF7细胞STAT4基因的表达水平(P<0.05),免疫印迹实验也证实经丹参酮ⅡA干预后MCF7细胞中STAT4蛋白含量增加,分子对接结果表明丹参酮ⅡA和STAT4蛋白能够形成稳定的结合构象。结论:STAT4基因在乳腺癌中表达较正常乳腺组织中偏低,且与乳腺癌患者的预后密切相关。丹参酮ⅡA能够上调MCF7细胞STAT4的基因和蛋白表达水平,该基因可能是丹参酮ⅡA干预乳腺癌的靶点之一。展开更多
基金supported in part by grants from National Natural Science Foundation of China(Grant Nos.82172635,82272686,and 82203628)the Natural Science Foundation of Tianjin(Grant Nos.23JCZDJC00200 and 21JCYBJC01000)the Tianjin Key Medical Discipline(Specialty)Construction Project(Grant No.TJYXZDXK-010A).
文摘Objective:This study aimed at exploring the effects of the epigenetic regulator,chidamide,on reprogramming the immunosuppressive tumor microenvironment in small cell lung cancer(SCLC),particularly the roles in macrophage polarization and angiogenesis.The therapeutic efficacy of combining chidamide with the anti-angiogenic agent,anlotinib,for refractory SCLC was also evaluated.Methods:RNA sequencing and functional validation were performed to assess chidamide’s effects on macrophages.Signal transducer and activator of transcription 4(STAT4)-mediated transcriptional activation of CCL2 was confirmed with ChIP-qPCR.The synergistic efficacy of chidamide in combination with anlotinib was tested in preclinical models.Results:Chidamide enhanced macrophage infiltration and induced macrophage polarization toward the anti-tumor M1 phenotype.Mechanistically,chidamide upregulated CCL2 via STAT4 transcriptional activation,thereby reshaping the tumor immune microenvironment(TIME).Combining chidamide with anlotinib synergistically suppressed tumor growth and remodeled the immunosuppressive TME in SCLC in vivo.Conclusions:Chidamide reshaped the SCLC TIME by activating STAT4/CCL2,thus driving M1 macrophage polarization and enhancing anti-tumor immunity.Our findings highlight coordinated TIME-targeted therapy as a translatable strategy to overcome therapeutic resistance in SCLC and provide a rationale for clinical trials examining epigenetic and anti-angiogenic therapeutics combinations.
文摘目的观察喘可治注射液(CKZ)对慢性阻塞性肺疾病(COPD)大鼠肺组织IL-12/IL-12R/STAT4信号通路的影响。方法 50只SD大鼠随机分为正常组,模型组,CKZ高、中、低剂量(CKZ-H、CHZ-M、CKZ-L)组。除正常组外,各组复制COPD大鼠模型,并给予相应药物治疗,观察CKZ对COPD大鼠血清和肺泡灌洗液(BALF)中γ-干扰素(IFN-γ)、白细胞介素12(IL-12)、白细胞介素12受体(IL-12R)以及肺组织信号转导子和转录激活子4(STAT4)m RNA表达的影响。结果喘可治注射液各剂量组肺组织病理形态均有不同程度的改善,且降低COPD大鼠血清和BALF中IFN-γ、IL-12、IL-12R(除CKZ-L组BALF中IL-12R外)水平(P<0.05,P<0.01),可抑制大鼠肺组织STAT4 m RNA的表达(P<0.01,P<0.05)。结论喘可治注射液能一定程度改善COPD大鼠肺组织病理损害,其作用机制可能与其抑制IFN-γ、IL-12、IL-12R表达,干扰IL-12/IL-12R/STAT4信号通路对靶基因的激活,下调STAT4 m RNA表达,进而调节Th1/Th2分化失衡有关。
文摘为了阐明水牛信号转导子和转录激活子4(signal transducer and activator of transcription 4,STAT4)基因在水牛卵泡、胚胎发生及泌乳调节过程中的作用及分子机制,本试验采用了3′-RACE克隆获得STAT4基因,并对其核苷酸序列和蛋白质序列进行了信息学分析,通过构建其真核表达载体并转染HEK293T细胞验证所构建载体的准确性。结果表明,水牛STAT4基因编码区长2 247bp,3′-UTR区长268bp,编码748个氨基酸。BLAST分析显示,水牛STAT4核苷酸序列与牛、山羊、绵羊、猪、马、犬和人相应序列的同源性分别为99%、99%、99%、95%、93%、93%和92%,系统进化树分析结果表明,STAT4基因在不同物种及进化的过程中具有高度保守性。蛋白质分析结果表明,STAT4蛋白呈弱酸性,无信号肽,定位于细胞质,存在STAT_int、STAT_alpha、STAT_bind和SH2_STAT4等结构域。microRNA靶标预测显示bta-miR-200a、bta-miR-2429和bta-miR-2410等为STAT4 3′-UTR潜在microRNAs。试验成功构建了水牛STAT4基因真核表达载体pEGFPN1-STAT4,转染HEK293T后产生较强的绿色荧光信号,表明能够形成STAT4-EGFP融合蛋白。
文摘目的:整理挖掘TCGA(the Cancer Genome Atlas)和GEO(gene expression omnibus)数据库中乳腺癌基因表达数据集的相关数据,分析STAT4基因在乳腺癌中的表达及临床意义,探讨丹参酮ⅡA对乳腺癌细胞中STAT4基因的干预效果。方法:下载TCGA数据库中的乳腺癌数据集(TCGA Breast Cancer(BRCA))和GEO数据库中有关乳腺癌和丹参酮ⅡA干预乳腺癌的数据集(GSE42586、GSE85871),对所获取的数据资料进行整理并作统计分析,通过免疫印迹实验和分子对接对数据分析结果进行验证。结果:对TCGA和GEO数据库中所有关于乳腺癌和正常组织乳腺的STAT4基因表达数据进行统计分析,结果发现乳腺癌组织中STAT4基因的表达明显低于正常乳腺组织,差异有统计学意义(P<0.05)。通过生存分析发现高表达STAT4基因的乳腺癌患者无病生存期明显好于低表达者,高表达患者的预后更好(P<0.05)。GSE85871数据集中有关丹参酮ⅡA作用于乳腺癌MCF7细胞的基因表达数据表明丹参酮ⅡA可以上调MCF7细胞STAT4基因的表达水平(P<0.05),免疫印迹实验也证实经丹参酮ⅡA干预后MCF7细胞中STAT4蛋白含量增加,分子对接结果表明丹参酮ⅡA和STAT4蛋白能够形成稳定的结合构象。结论:STAT4基因在乳腺癌中表达较正常乳腺组织中偏低,且与乳腺癌患者的预后密切相关。丹参酮ⅡA能够上调MCF7细胞STAT4的基因和蛋白表达水平,该基因可能是丹参酮ⅡA干预乳腺癌的靶点之一。
