特异性敲除调节性 T 细胞中的 Notch2 显著抑制小鼠头颈部鳞状细胞癌的生长和增殖。
Specific knockout of Notch2 in Treg cells significantly inhibits the growth and proliferation of head and neck squamous cell carcinoma in mice.
机构信息
Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, 238 Jie-Fang Road, Wuhan, Hubei 430060, PR China.
Department of Otolaryngology Head and Neck Surgery, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, PR China.
出版信息
Int Immunopharmacol. 2023 Oct;123:110705. doi: 10.1016/j.intimp.2023.110705. Epub 2023 Jul 29.
OBJECTIVE
To investigate the effect of Notch2 gene knockout in Treg cells on head and neck squamous cell carcinoma (HNSCC) in mice.
METHODS
A mouse model of HNSCC was constructed. Flow cytometry and immunofluorescence were used to examine the numbers of related immune cells and programmed cell death in tumor cells in the spleen and tumor microenvironment of mice. Western blotting was used to measure the expression of related proteins in tumor tissues.
RESULTS
The tumor volume of regulatory T (Treg) cell-specific Notch2-knockout mice (experimental group) was significantly smaller than that of control mice (control group) (P < 0.05). Compared with those in the control group, the number of Treg cells and the expression of Ki67 in Treg cells in the spleen and tumor tissue were significantly decreased in the experimental group, while the numbers of CD45+ hematopoietic cells, CD4+ T cells, CD8+ T cells, T helper 1 (Th1) cells, CD11b+ cells (macrophages), and CD11b+CD11c+ cells (dendritic cells) and the expression of Ki67 in CD4+ T cells and CD8+ T cells were significantly increased (P < 0.05). There was no significant difference in the number of Th2 cells between the two groups (P > 0.05). Immunofluorescence analysis showed that the numbers of CD4+ T cells and CD8+ T cells in the tumor tissue in the experimental group were significantly higher than those in the control group (P < 0.05). Compared with that in the control group, programmed cell death in the experimental group was significantly increased (P < 0.05). Moreover, the expression levels of NLRP3, Caspase-1 and GSDMD in the tumor tissues of the experimental group were higher than those in the control group (P < 0.01), while the expression levels of BCL2, Bax, ATG5, LC3 and p62 were not significantly different (P > 0.05).
CONCLUSIONS
Specific knockout of the Notch2 gene in Treg cells significantly decreases the function of Treg cells, inhibits the growth of HNSCC and improves the immune microenvironment in mice, thus effectively treating HNSCC.
目的
研究 Notch2 基因敲除对调节性 T 细胞（Treg）对小鼠头颈部鳞状细胞癌（HNSCC）的影响。
方法
构建了小鼠 HNSCC 模型。通过流式细胞术和免疫荧光法检测小鼠脾脏和肿瘤微环境中相关免疫细胞和肿瘤细胞程序性细胞死亡的数量。Western blot 法检测肿瘤组织中相关蛋白的表达。
结果
与对照组（对照组）相比，Treg 细胞特异性 Notch2 基因敲除小鼠（实验组）的肿瘤体积明显减小（P<0.05）。与对照组相比，实验组脾脏和肿瘤组织中 Treg 细胞数量和 Ki67 在 Treg 细胞中的表达明显减少，而 CD45+造血细胞、CD4+T 细胞、CD8+T 细胞、Th1 细胞、CD11b+细胞（巨噬细胞）和 CD11b+CD11c+细胞（树突状细胞）的数量以及 CD4+T 细胞和 CD8+T 细胞中 Ki67 的表达明显增加（P<0.05）。两组间 Th2 细胞数量无明显差异（P>0.05）。免疫荧光分析显示，实验组肿瘤组织中 CD4+T 细胞和 CD8+T 细胞数量明显高于对照组（P<0.05）。与对照组相比，实验组细胞程序性死亡明显增加（P<0.05）。此外，实验组肿瘤组织中 NLRP3、Caspase-1 和 GSDMD 的表达水平高于对照组（P<0.01），而 BCL2、Bax、ATG5、LC3 和 p62 的表达水平无明显差异（P>0.05）。
结论
Treg 细胞中 Notch2 基因的特异性敲除显著降低了 Treg 细胞的功能，抑制了 HNSCC 的生长，改善了小鼠的免疫微环境，从而有效治疗 HNSCC。
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