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

doi:10.13701/j.cnki.kqyxyj.2023.08.005

口腔医学研究 ›› 2023, Vol. 39 ›› Issue (8): 684-689.DOI: 10.13701/j.cnki.kqyxyj.2023.08.005

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

李易玮, 孙雅楠, 潘军臣, 胡雅英, 张钰莹, 马纪源, 张佳莉^*

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

收稿日期:2023-01-12 发布日期:2023-08-17
通讯作者: *张佳莉,E-mail:jiali_zhang@whu.edu.cn
作者简介:李易玮(1997~ ),女,湖北武汉人,硕士在读,主要从事口腔肿瘤基础研究。
基金资助:
国家自然科学基金(编号:82273201、81972552)

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

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

Received:2023-01-12 Published:2023-08-17

摘要/Abstract

摘要： 目的: 探究BarH-样同源框蛋白2(BarH-like homeobox 2,Barx2)通过促进MHC Ⅱ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。

关键词: BarH-样同源框蛋白2, MHC-Ⅱ, 口腔鳞状细胞癌, 肿瘤免疫

Abstract: Objective: To explore the mechanism of BarH-like homeobox 2 (Barx2) killing tumor cells by promoting MHC Class Ⅱ molecules. Methods: The expression levels of CⅡTA/MHC-Ⅱ related molecules in Barx2 overexpressed OSCC cell lines were analyzed by high-throughput sequencing, qPCR, and Western Blot. The combination of Barx2 and CⅡTA pⅢ promoter 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-Ⅱ signaling 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 pⅢ promoter. 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-Ⅱ signal axis, induce the expression of HLA-DR-dominated MHC-Ⅱ class molecules in tumor cells, and further activate CD4⁺T and CD8⁺T cells to play a role in killing tumor cells.

Key words: BarH-like homeobox 2, MHC-Ⅱ, oral squamous cell carcinoma, immunity to tumors

李易玮, 孙雅楠, 潘军臣, 胡雅英, 张钰莹, 马纪源, 张佳莉. 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[J]. 口腔医学研究, 2023, 39(8): 684-689.

LI Yiwei, SUN Yanan, PAN Junchen, HU Yaying, ZHANG Yuying, MA Jiyuan, ZHANG Jiali. Barx2 Drives Tumor-specific MHC-Ⅱ Signaling to Induce CD4⁺T/CD8⁺T Cell Activation in Oral Squamous Cell Carcinoma[J]. Journal of Oral Science Research, 2023, 39(8): 684-689.

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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-Ⅱ Signaling to Induce CD4⁺T/CD8⁺T Cell Activation in Oral Squamous Cell Carcinoma

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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-Ⅱ Signaling to Induce CD4+T/CD8+T Cell Activation in Oral Squamous Cell Carcinoma

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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-Ⅱ Signaling to Induce CD4⁺T/CD8⁺T Cell Activation in Oral Squamous Cell Carcinoma