小鼠舌白斑的转录组学特征和信号通路改变及白介素17A单抗治疗的实验研究  

Experimental study on the transcriptomic characteristics and signaling pathway alteration in mouse tongue leukoplakia and effect of anti-mouse IL-17A monoclonal antibody on leukoplakia

作　　者：韩馨怡 张倩倩 王子瑞 周曾同[1] 唐国瑶[1,2] 施琳俊 沈雪敏[1] HAN Xin-yi;ZHANG Qian-qian;WANG Zi-rui;ZHOU Zeng-tong;TANG Guo-yao;SHI Lin-jun;SHEN Xue-min(The Department of Oral Medicine,Shanghai Ninth People's Hospital,Shanghai JiaoTong University School of Medicine·Department of Oral Mucosa,College of Stomatology,Shanghai JiaoTong University,National Center for Stomatology,National Clinical Research Center for Oral Diseases,Shanghai Key Laboratory of Stomatology,Shanghai Research Institute of Stomatology,Shanghai 20001l,China;Department of Stomatology,Shanghai Xin Hua Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China)

机构地区：[1]上海交通大学医学院附属第九人民医院·口腔黏膜病科,上海交通大学口腔医学院,国家口腔医学中心,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海2200011 [2]上海交通大学医学院附属新华医院,上海200092

出　　处：《临床口腔医学杂志》2024年第4期195-199,共5页Journal of Clinical Stomatology

基　　金：国家自然科学基金项目(82370954,82170952);上海市卫生健康委员会2020年度综合医院中西医结合专项立项一般项目(ZHYY-ZXYJHZX-202016);上海交通大学医学院附属第九人民医院医学研究培育基金(JYZZ182);上海交通大学医学院附属第九人民医院研究型学科(KQYJXK2021);上海市教委口腔高峰学科——临床医学“研究型医师”(双百人)(2022sbr-slj);科技部国家重点研发计划(SQ2022YFC2400066)。

摘　　要：目的:探索口腔白斑形成及进展过程中的转录组学特征和信号通路改变以及白介素17A(interleukin-17A,IL-17A)单抗对小鼠舌白斑的治疗作用。方法:分析舌白斑患者同时期正常黏膜组织(NM)、白斑伴上皮异常增生组织(OL)和小鼠正常舌黏膜组织(mNM)、4NQO诱导的小鼠舌白斑伴异常增生组织(mOL)的转录组学特征;4NQO诱导12周小鼠舌白斑形成后将小鼠分为实验组(腹腔注射IL-17A单克隆抗体)和对照组(腹腔注射IgG1同型对照抗体)两组,其中单抗的剂量为10 mg/kg/w。注射4周后取材并通过苏木精-伊红染色(HE染色)和Ki67免疫组织化学染色(IHC)的方法对小鼠舌进行组织病理学评价;体外IL-17A刺激人口腔白斑细胞系(DOK、Leuk1),并用流式细胞术分析其细胞周期。结果:综合分析人和小鼠的转录组测序结果,显示IL-17信号通路在白斑组织中显著上调。小鼠舌白斑用药4周后,实验组小鼠舌80%为轻到中度异常增生,20%为重度异常增生,而对照组中40%为轻到中度异常增生,40%为重度异常增生,20%为浸润癌。Ki67结果显示对照组小鼠舌上皮细胞中处于增殖时期的细胞比例显著多于对照组(P<0.05)。流式细胞术显示,相较对照组,IL-17A显著导致白斑细胞更多集中在S-G2-M期(P<0.05)。结论:人及小鼠舌白斑形成及进展过程中IL-17信号通路显著上调;IL-17A单抗对4NQO诱导的小鼠舌白斑有潜在的治疗作用;IL-17A会促进人白斑细胞的增殖。Objective:To explore the transcriptomic characteristics and signal pathway changes of mouse tongue leukoplakia formation and malignant progression,as well as the therapeutic effect of interleukin-17A(IL-17A)monoclonal antibody on mouse tongue leukoplakia.Methods:RNA-sequence was performed on human normal tongue mucosa tissues(NM),tongue leukoplakia(OL)tissues with epithelial dysplasia from the same patient,normal mucosa tissues,and leukoplakia tissues(mOL)from the tongue of 4NQO-induced mice.After 12 weeks of 4NQO induction,the mice were divided into two groups:experimental group(intraperitoneal injection of anti-mouse IL-17A monoclonal antibody)and control group(intraperitoneal injection of IgG1 isotype control antibody),the dosage of monoclonal antibody was 10 mg/kg/w.Tissue samples were collected after four weeks'injection.And the histopathology of the mouse tongues were performed by hematoxylin-eosin staining(HE staining)and immunohistochemistry staining(IHC)of Ki67.In vitro,the leukoplakia cell lines(DOK,Leuk1)were stimulated with recombinant human IL-17A respectively and cell cycle was analyzed by flow cytometry.Results:Comprehensive analysis of the transcriptomic sequencing results of humans and mice showed that the IL-17 signaling pathway was significantly upregulated during leukoplakia formation.The histopathological evaluation results after four weeks of Anti-mouse IL-17A monoclonal antibody treatment showed that 80%of the mouse tongues in the experimental group had mild to moderate epithelial dysplasia,while 20%had severe epithelial dysplasia.In the control group,40%had mild to moderate epithelial dysplasia,40%had severe epithelial dysplasia,but 20%had infiltrating cancer.The results of Ki67 showed that the proportion of cells in the proliferation stage in the control group was significantly higher than that in the control group(P<0.05).And flow cytometry showed that IL-17A stimulation led to more concentration of cells in the S-G2-M phase(P<0.05).Conclusion:The IL-17 signaling pathway is significantly

关 键 词：转录组测序 IL-17信号通路 4NQO小鼠口腔白斑模型 IL-17A单抗 

分 类 号：R781.5[医药卫生—口腔医学]