牙龈卟啉单胞菌通过CCL20-CCR6轴构建口腔鳞癌免疫抑制性微环境的实验研究

doi:10.19438/j.cjoms.2025.06.002

摘要/Abstract

摘要： 目的：探讨牙龈卟啉单胞菌（Porphyromonas gingivalis,P.gingivalis）通过CCL20-CCR6通路形成口腔鳞癌（oral squamous cell carcinoma,OSCC）免疫抑制性微环境的分子机制。方法：体外培养鼠源性鳞癌细胞系SCC7和P.gingivalis,建立C57BL/6小鼠肿瘤模型。基于对P.gingivalis的感染情况和CCL20-CCR6轴的抑制,将C57BL/6小鼠分为对照组（SCC7）、实验组（SCC7+P.g）和干预组（SCC7+P.g+CCR6 antagonist）,大体观察不同分组肿瘤质量、体积的变化,采用免疫组织化学检测肿瘤组织CD4、FOXP3、PD-1、CTLA-4、LAG3、TIM3、TIGIT的表达,采用ELISA法检测血清TGF-β的含量。结果：与对照组相比,实验组P.gingivalis感染显著促进了OSCC荷瘤小鼠肿瘤组织的体积增长（P<0.05）,肿瘤组织免疫微环境中CD4、Foxp3、CTLA-4、LAG3、TIM3、TIGIT、TGF-β表达显著增加（P<0.05）,PD-1表达无显著差异（P>0.05）。干预组与实验组相比,小鼠肿瘤组织体积显著缩小（P<0.05）,相关免疫抑制性分子和免疫检查点CD4、Foxp3、PD-1、CTLA-4、LAG3、TIGIT、TGF-β表达显著降低（P<0.05）,TIM3表达无统计学差异（P>0.05）。结论：P.gingivalis通过CCL20-CCR6轴招募更多的Treg细胞,调节肿瘤相关抑制性细胞因子和免疫抑制性检查点的表达,进而促进OSCC的免疫抑制性微环境形成。

关键词: 牙龈卟啉单胞菌, 口腔鳞癌, 免疫抑制性微环境, 调节性T细胞, CCL20-CCR6

Abstract: PURPOSE: To explore the molecular mechanism by which Porphyromonas gingivalis(P. gingivalis) forms an immunosuppressive microenvironment in oral squamous cell carcinoma(OSCC) through the CCL20-CCR6 pathway. METHODS: Mouse-derived squamous cell carcinoma cell line SCC7 and P. gingivalis were cultured in vitro, and C57BL/6 mice were used to establish a tumor mouse model. Based on the infection status of P.gingivalis and the inhibition of the CCL20-CCR6 axis, C57BL/6 mice were divided into control group(SCC7), experimental group(SCC7+P.g), and intervention group(SCC7+P.g+CCR6 antagonist). The changes in tumor mass and volume among different groups were observed macroscopically, immunohistochemistry was used to detect CD4, FOXP3, PD-1, CTLA-4, LAG3, TIM3 and TIGIT in tumor tissues, ELISA was applied to measure the serum TGF-β levels. RESULTS: Compared to the control group, P.gingivalis infection in the experimental group significantly promoted the tumor tissue volume in OSCC-bearing mice(P<0.05), and the expression of CD4, Foxp3, CTLA-4, LAG3, TIM3, TIGIT, and TGF-β in the tumor tissue immunological microenvironment was significantly increased (P<0.05), while PD-1 expression showed no significant difference (P>0.05). The intervention group showed a reduction in tumor tissue volume compared to the experimental group (P<0.05), the expression of relevant immunosuppressive molecules and immune checkpoints CD4, Foxp3, PD-1, CTLA-4, LAG3, TIGIT, and TGF-β decreased(P<0.05), while TIM3 expression showed no significant difference(P>0.05). CONCLUSIONS: P.gingivalis recruits more Treg cells through the CCL20-CCR6 axis, regulates the expression of tumor-associated suppressive cytokines and immunosuppressive checkpoints, thereby promoting the formation of an immunosuppressive microenvironment in OSCC.

Key words: Porphyromonas gingivalis, Oral squamous cell carcinoma, Immunosuppressive microenvironment, Regulatory T cells, CCL20-CCR6

中图分类号:

R739.81

乃吉拜·莫敏, 许立明, 张一博, 李娇阳, 刘朗清, 凌彬. 牙龈卟啉单胞菌通过CCL20-CCR6轴构建口腔鳞癌免疫抑制性微环境的实验研究[J]. 中国口腔颌面外科杂志, 2025, 23(6): 546-552.

Naijibai·Momin, Xu Liming, Zhang Yibo, Li Jiaoyang, Liu Langqing, Ling Bin. Mechanism of Porphyromonas gingivalis in establishing an immunosuppressive microenvironment in oral squamous cell carcinoma via the CCL20-CCR6 axis[J]. China J Oral Maxillofac Surg, 2025, 23(6): 546-552.

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