Barx2驱动口腔鳞状细胞癌肿瘤特异性MHC-Ⅱ类信号诱导CD4^(+)T/CD8^(+)T细胞活化  

Barx2 Drives Tumor-specific MHC-ⅡSignaling to Induce CD4^(+)T/CD8^(+)T Cell Activation in Oral Squamous Cell Carcinoma

作　　者：李易玮 孙雅楠[1] 潘军臣 胡雅英 张钰莹 马纪源 张佳莉[1] LI Yiwei;SUN Yanan;PAN Junchen;HU Yaying;ZHANG Yuying;MA Jiyuan;ZHANG Jiali(State Key Laboratory of Oral&Maxillofacial Reconstruction and Regeneration,Key Laboratory of Oral Biomedicine Ministry of Education,Hubei Key Laboratory of Stomatology,Department of Oral Pathology,School&Hospital of Stomatology,Wuhan University,Wuhan 430079,China)

机构地区：[1]口颌系统重建与再生全国重点实验室,口腔生物医学教育部重点实验室,口腔医学湖北省重点实验室,武汉大学口腔医(学)院口腔病理科,湖北武汉430079

出　　处：《口腔医学研究》2023年第8期684-689,共6页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:82273201、81972552)。

摘　　要：目的:探究BarH-样同源框蛋白2(BarH-like homeobox 2,Barx2)通过促进MHCⅡ类信号通路杀伤肿瘤细胞的机制。方法:(1)通过高通量测序,qPCR和Western Blot实验分析Barx2过表达的OSCC细胞系中CⅡTA/MHC-Ⅱ信号轴相关分子的表达量;(2)双荧光素酶实验验证Barx2与CⅡTA pⅢ启动子的结合;(3)提取人外周血单核细胞(PBMC),分别与对照组和Barx2过表达组肿瘤细胞共培养。共培养后,结晶紫染色检测肿瘤细胞的数量;应用CFSE染色标记及流式细胞学实验分析PBMC中CD4^(+)T细胞和CD8^(+)T细胞增殖活性。结果:(1)高通量转录组测序结果结合GSEA-基因集富集分析和KEGG信号通路分析发现,过表达Barx2上调了抗原加工与提呈通路MHC-Ⅱ信号分子相关因子;在Barx2过表达细胞系中,qPCR及Western blot进一步验证了上述基因表达的显著上调;(2)Barx2能直接与CⅡTA pⅢ启动子上游结合,促进主要表达于淋巴细胞的CⅡTA转录本的转录。(3)肿瘤细胞与PBMC共培养后,Barx2过表达肿瘤细胞的增殖被显著抑制;过表达Barx2的肿瘤细胞显著促进了CD4^(+)T和CD8^(+)T细胞的增殖活性。结论:Barx2通过驱动CⅡTA/MHC-Ⅱ信号轴活化,促进肿瘤细胞表达以HLA-DR为主的MHC-Ⅱ类分子,从而活化CD4^(+)T细胞和CD8^(+)T细胞,发挥杀伤肿瘤细胞的作用。Objective:To explore the mechanism of BarH-like homeobox 2(Barx2)killing tumor cells by promoting MHC ClassⅡmolecules.Methods:The expression levels of CⅡTA/MHC-Ⅱrelated molecules in Barx2 overexpressed OSCC cell lines were analyzed by high-throughput sequencing,qPCR,and Western Blot.The combination of Barx2 and CⅡTA pⅢpromoter was verified by double luciferase assay.Human peripheral blood mononuclear cells(PBMC)were extracted and co-cultured with tumor cells in the control group and Barx2 overexpression group,and the number of tumor cells was detected by crystal violet staining.The proliferation activity of CD4^(+)T cells and CD8^(+)T cells in PBMC was analyzed by flow cytometry after labeling PBMC with CFSE.Results:High-throughput transcriptome sequencing results combined with GSEA gene set enrichment analysis and KEGG signaling pathway analysis showed that overexpression of Barx2 enhanced the up-regulation of factors related to antigen processing and presentation pathway and MHC-Ⅱsignaling molecules.In Barx2 overexpressed cell lines,qPCR and Western blot further confirmed the significant upregulation of the above genes.Barx2 could directly bind upstream to CⅡTA pⅢpromoter.After co-culture with PBMC,the number of tumor cells overexpressing Barx2 was significantly less than that in the control group.Overexpression of Barx2 tumor cells promoted the proliferation of CD4^(+)T and CD8^(+)T cells.Conclusion:Barx2 can drive the activation of CⅡTA/MHC-Ⅱsignal axis,induce the expression of HLA-DR-dominated MHC-Ⅱclass molecules in tumor cells,and further activate CD4^(+)T and CD8^(+)T cells to play a role in killing tumor cells.

关 键 词：BarH-样同源框蛋白2 MHC-Ⅱ 口腔鳞状细胞癌 肿瘤免疫 

分 类 号：R739.8[医药卫生—肿瘤]