Background:Evading immune surveillance is necessary for tumor metastasis.Thus,there is an urgent need to better understand the interaction between metastasis and mechanisms of tumor immune evasion.In this study,we aim...Background:Evading immune surveillance is necessary for tumor metastasis.Thus,there is an urgent need to better understand the interaction between metastasis and mechanisms of tumor immune evasion.In this study,we aimed to clarify a novel mechanism that link tumor metastasis and immunosuppression in the development of esophageal squamous cell carcinoma(ESCC).Methods:The expression of melanoma-associated antigen C3(MAGE-C3)was detected using immunohistochemistry.Transwell assays were used to evaluate the migration and invasion ability of esophageal squamous cell carcinoma(ESCC)cells.Metastasis assays in mice were used to evaluate metastatic ability in vivo.Lymphocyte-mediated cytotoxicity assays were performed to visualize the immune suppression function on tumor cells.RNA sequencing was performed to identify differentially expressed genes between MAGE-C3 overexpressing ESCC cells and control cells.Gene ontology(GO)enrichment analyses was performed to identify the most altered pathways influenced by MAGE-C3.The activation of the interferon-γ(IFN-γ)pathway was analyzed using Western blotting,GAS luciferase reporter assays,immunofluorescence,and flow cytometry.The role of MAGE-C3 in the IFN-γpathway was determined by Western blotting and immunoprecipitation.Furthermore,immunohistochemistry and flow cytometry analysis monitored the changes of infiltrated T cell populations in murine lung metastases.Results:MAGE-C3 was overexpressed in ESCC tissues.High expression of MAGE-C3 had a significant association with the risk of lymphatic metastasis and poor survival in patients with ESCC.Functional experiments revealed that MAGE-C3 promoted tumor metastasis by activating the epithelial-mesenchymal transition(EMT).MAGE-C3 repressed antitumor immunity and regulated cytokine secretion of T cells,implying an immunosuppressive function.Mechanistically,MAGE-C3 facilitated IFN-γsignaling and upregulated programmed cell death ligand 1(PDŋL1)by binding with IFN-γreceptor 1(IFNGR1)and strengthening the interaction between IFNGR1展开更多
目的探讨1例疑似孟德尔遗传易感分枝杆菌病（Mendelian susceptibility to mycobacterial disease,MSMD）患儿的临床特征,检测IL-12/23-IFN-γ通路的完整性,并进行IFN-γ受体1（IFNGR1）基因分析。方法根据患儿的临床表现及常规的免疫...目的探讨1例疑似孟德尔遗传易感分枝杆菌病（Mendelian susceptibility to mycobacterial disease,MSMD）患儿的临床特征,检测IL-12/23-IFN-γ通路的完整性,并进行IFN-γ受体1（IFNGR1）基因分析。方法根据患儿的临床表现及常规的免疫学筛查试验排除常见原发性免疫缺陷病,Q-RT-PCR在mRNA水平检测患儿及健康对照者IL-12/23-IFN-γ通路的完整性,通过PCR及RT-PCR分别对患儿及其父母的IFNGR1基因进行扩增并测序。结果患儿有播散性卡介苗（BCG）病及全身多系统损害的表现。患儿全血经BCG和IFN-γ共同刺激48 h后IL-12B的表达水平与单独BCG刺激后比较明显降低（P〈0.05）;基因测序发现患儿IFNGR1第6外显子818-821位杂合缺失4个碱基（c.818-821 del TTAA）,使第276位脯氨酸突变为终止密码（p.Asn274fsX276）,父母此位点正常。结论 MSMD患儿临床特征为BCG病及全身多系统损害。Q-RT-PCR检测IL-12/23-IFN-γ通路完整性是一种有效的筛查MSMD的手段;IFNGR1突变是引起本例患儿发生上述临床特征的根本原因。展开更多
基金This study was supported by grants from the National Natural Science Foundation of China(81988101,81830086,82003007 and 81802780)CAMS Innovation Fund for Medical Sciences(2019-I2M-5-081)+3 种基金Guangdong Basic and Applied Basic Research Foundation(2019B030302012)the Funding by Major Program of Shenzhen Bay Laboratory(S201101004)Project funded by China Postdoctoral Science Foundation(2019M6603068)Science Foundation of Peking University Cancer Hospital(2020-16).
文摘Background:Evading immune surveillance is necessary for tumor metastasis.Thus,there is an urgent need to better understand the interaction between metastasis and mechanisms of tumor immune evasion.In this study,we aimed to clarify a novel mechanism that link tumor metastasis and immunosuppression in the development of esophageal squamous cell carcinoma(ESCC).Methods:The expression of melanoma-associated antigen C3(MAGE-C3)was detected using immunohistochemistry.Transwell assays were used to evaluate the migration and invasion ability of esophageal squamous cell carcinoma(ESCC)cells.Metastasis assays in mice were used to evaluate metastatic ability in vivo.Lymphocyte-mediated cytotoxicity assays were performed to visualize the immune suppression function on tumor cells.RNA sequencing was performed to identify differentially expressed genes between MAGE-C3 overexpressing ESCC cells and control cells.Gene ontology(GO)enrichment analyses was performed to identify the most altered pathways influenced by MAGE-C3.The activation of the interferon-γ(IFN-γ)pathway was analyzed using Western blotting,GAS luciferase reporter assays,immunofluorescence,and flow cytometry.The role of MAGE-C3 in the IFN-γpathway was determined by Western blotting and immunoprecipitation.Furthermore,immunohistochemistry and flow cytometry analysis monitored the changes of infiltrated T cell populations in murine lung metastases.Results:MAGE-C3 was overexpressed in ESCC tissues.High expression of MAGE-C3 had a significant association with the risk of lymphatic metastasis and poor survival in patients with ESCC.Functional experiments revealed that MAGE-C3 promoted tumor metastasis by activating the epithelial-mesenchymal transition(EMT).MAGE-C3 repressed antitumor immunity and regulated cytokine secretion of T cells,implying an immunosuppressive function.Mechanistically,MAGE-C3 facilitated IFN-γsignaling and upregulated programmed cell death ligand 1(PDŋL1)by binding with IFN-γreceptor 1(IFNGR1)and strengthening the interaction between IFNGR1
文摘目的探讨1例疑似孟德尔遗传易感分枝杆菌病（Mendelian susceptibility to mycobacterial disease,MSMD）患儿的临床特征,检测IL-12/23-IFN-γ通路的完整性,并进行IFN-γ受体1（IFNGR1）基因分析。方法根据患儿的临床表现及常规的免疫学筛查试验排除常见原发性免疫缺陷病,Q-RT-PCR在mRNA水平检测患儿及健康对照者IL-12/23-IFN-γ通路的完整性,通过PCR及RT-PCR分别对患儿及其父母的IFNGR1基因进行扩增并测序。结果患儿有播散性卡介苗（BCG）病及全身多系统损害的表现。患儿全血经BCG和IFN-γ共同刺激48 h后IL-12B的表达水平与单独BCG刺激后比较明显降低（P〈0.05）;基因测序发现患儿IFNGR1第6外显子818-821位杂合缺失4个碱基（c.818-821 del TTAA）,使第276位脯氨酸突变为终止密码（p.Asn274fsX276）,父母此位点正常。结论 MSMD患儿临床特征为BCG病及全身多系统损害。Q-RT-PCR检测IL-12/23-IFN-γ通路完整性是一种有效的筛查MSMD的手段;IFNGR1突变是引起本例患儿发生上述临床特征的根本原因。
