牙龈卟啉单胞菌诱发炎症微环境促进食管鳞状细胞癌发生  

Porphyromonas gingivalis promotes the occurrence of esophageal squamous cell carcinoma via an inflammatory microenvironment

作　　者：许海军 齐义军 伍当柔 刘其伟 陈攀[1] 李孟祥 焦叶林 阮豪杰 李智涛[1] 高社干[1] Xu Haijun;Qi Yijun;Wu Dangrou;Liu Qiwei;Chen Pan;Li Mengxiang;Jiao Yelin;Ruan Haojie;Li Zhitao;Gao Shegan(State Key Laboratory of Esophageal Cancer Prevention&Treatment,Henan Key Laboratory of Microbiome and Esophageal Cancer Prevention and Treatment,Henan Key Laboratory of Cancer Epigenetics,Cancer Hospital,The First Affiliated Hospital,College of Clinical Medicine of Henan University of Science and Technology,Luoyang 471003,China;Key Laboratory of Cell Behavior,Medical School of Xuchang University,Xuchang 461000,China;Department of Mathematics and Physics,Luoyang Institute of Science and Technology,Luoyang 471023,China;Department of Pathology,Luoyang First People′s Hospital,Luoyang 471000,China)

机构地区：[1]河南科技大学临床医学院、河南科技大学第一附属医院肿瘤医院,省部共建食管癌防治国家重点实验室,河南省微生态与食管癌防治重点实验室,河南省肿瘤表观遗传重点实验室,洛阳471003 [2]许昌学院医学院细胞行为学重点实验室,许昌461000 [3]洛阳理工学院数学与物理教学部,洛阳471023 [4]洛阳市第一人民医院病理科,洛阳471000

出　　处：《中华肿瘤杂志》2024年第8期746-754,共9页Chinese Journal of Oncology

基　　金：国家自然科学基金(81872037,81972571);河南省教育厅高校重点科研项目(24A310015)。

摘　　要：目的探讨牙龈卟啉单胞菌诱发食管炎症微环境在小鼠食管鳞状细胞癌发生过程中的作用。方法采用数字表法随机将180只C57BL/6小鼠分为对照组、牙龈卟啉单胞菌组、4-硝基喹啉-1-氧化物(4NQO)组、4NQO+牙龈卟啉单胞菌组、4NQO+牙龈卟啉单胞菌+塞来昔布组和4NQO+牙龈卟啉单胞菌+抗生素(甲硝唑、新霉素、氨苄青霉素和万古霉素,ABC)组,每组30只。给予ABC饮水2周,之后8周,对照组和牙龈卟啉单胞菌组小鼠饮用纯水,其余4组给予含30μg/ml 4NQO的饮水。第11~12周,牙龈卟啉单胞菌组、4NQO+牙龈卟啉单胞菌组、4NQO+牙龈卟啉单胞菌+塞来昔布组和4NQO+牙龈卟啉单胞菌+ABC组小鼠行上颌第2磨牙结扎;第11~34周,每周3次口腔感染牙龈卟啉单胞菌。第13~34周,4NQO+牙龈卟啉单胞菌+塞来昔布组和4NQO+牙龈卟啉单胞菌+ABC组小鼠分别接受塞来昔布和ABC处理。34周后处死小鼠,观察小鼠食管黏膜大体和形态学变化,采用实时荧光定量聚合酶链反应(RT-qPCR)和免疫组化检测小鼠食管组织中炎症和肿瘤相关分子表达。结果34周时,4NQO单独处理未能显著增加小鼠食管黏膜乳头状增生病灶数、病变面积和食管壁厚度,单纯性增生病灶数(中位数为1.00个,P<0.05)和轻、中度不典型增生病灶数(中位数为2.00个,P<0.01)显著增加,小鼠食管组织中白细胞介素6(IL-6)、IL-1β、肿瘤坏死因子α(TNF-α)、c-myc mRNA的表达量[分别为8.35(3.45,8.99)、6.90(2.01,9.72)、12.04(3.31,14.08)、2.21(1.80,3.04),均P<0.05]和磷酸化信号转导和转录激活因子3(pSTAT3)、Ki-67、磷酸化组蛋白2A变异体(pH2AX)蛋白的表达明显高于对照组。4NQO+牙龈卟啉单胞菌组小鼠食管黏膜病变显著,乳头状增生病灶数(中位数为2.00个)、病变面积(中位数为2.51 mm^(2))和食管壁厚度(中位数为172.52μm)最大,且均与对照组差异有统计学意义(均P<0.01),单纯性增生病灶数(中位数为1.00个,P<0.05)和轻Objective To investigate the role of an inflammatory microenvironment induced by Porphyromonasgingivalis(P.gingivalis)in the occurrence of esophageal squamous cell carcinoma(ESCC)in mice.Methods A total of 180 C57BL/6 mice were randomly divided into 6 groups,i.e.control group,P.gingivalis group,4NQO group,4NQO+P.gingivalis group,4NQO+P.gingivalis+celecoxib group,and 4NQO+P.gingivalis+antibiotic cocktail(ABC,including metronidazole,neomycin,ampicillin,and vancomycin)group,with 30 mice in each group,using the random number table.All mice were normalized by treatment with ABC in drinking water for 2 weeks.In the following 2 weeks,the mice in the control group and the P.gingivalis group were given drinking water,while the other 4 groups were treated with 30μg/ml 4NQO in the drinking water.In weeks 11-12,the mice in the P.gingivalis group,the 4NQO+P.gingivalis group,the 4NQO+P.gingivalis+celecoxib group,and the 4NQO+P.gingivalis+ABC group were subjected to ligation of the second molar in oral cavity followed by oral P.gingivalis infection thrice weekly for 24 weeks in weeks 11-34.In weeks 13-34,the mice in 4NQO+P.gingivalis+celecoxib group and 4NQO+P.gingivalis+ABC group were administered with celecoxib and ABC for 22 weeks,respectively.At the end of 34 weeks,gross and microscopic alterations were examined followed by RT-qPCR and immunohistochemistry to examine the expression profiles of inflammatory-and tumor-molecules in esophagi of mice.Results At 34 weeks,4NQO treatment alone did not affect the foci of papillary hyperproliferation,diseased area,and the thickness of the esophageal wall,but significantly enhanced the foci of hyperproliferation(median 1.00,P<0.05)and mild/moderate dysplasia(median 2.00,P<0.01).In addition,the expression levels of IL-6[8.35(3.45,8.99)],IL-1β[6.90(2.01,9.72)],TNF-α[12.04(3.31,14.08)],c-myc[2.21(1.80,3.04)],pSTAT3,Ki-67,and pH2AX were higher than those in the control group.The pathological changes of the esophageal mucosa were significantly more overt in the 4NQO+P.gingivalis group i

关 键 词：食管鳞状细胞癌 牙龈卟啉单胞菌 炎症微环境 4-硝基喹啉-1-氧化物 白细胞介素6 信号转导和转录激活因子3 环氧合酶2 组蛋白2A变异体 

分 类 号：R735.1[医药卫生—肿瘤]