舌黏膜癌变基因组甲基化分析  

Global analysis of DNA methylation changes during experimented lingual carcinogenesis

作　　者：刘华[1] 岳万远 邵帅 孙家平 杨莹[2] 代晓明[3] Liu Hua;Yue Wanyuan;Shao Shuai;Sun Jiaping;Yang Ying;Dai Xiaoming(Dept.of Oral and Maxillofacial Surgery,Affiliated Hospital of Yunnan University,Kunming 650021,China;Dept.of Pathology,First Affiliated Hospital of Kunming Medical University,Kunming 650031,China;Maxillofacial Service of Department of Plastic Surgery,First Affiliated Hospital of Kunming Medical University,Kunming 650031,China)

机构地区：[1]云南大学附属医院口腔颌面外科,昆明650021 [2]昆明医科大学第一附属医院病理科,昆明650031 [3]昆明医科大学第一附属医院整形外科颌面组,昆明650031

出　　处：《华西口腔医学杂志》2024年第3期319-328,共10页West China Journal of Stomatology

基　　金：云南省科技厅-昆医联合专项[202001AY070001-253,2018FE001(-216)]。

摘　　要：目的研究舌黏膜癌变过程中基因组甲基化特征,探讨舌癌中DNA甲基化的规律。方法用50 mg/L的4-硝基喹啉-1-氧化物(4NQO)饮水诱导C57BL/6J小鼠舌黏膜癌变,分别取第0、12、28周的舌黏膜(分别代表正常、癌前病变和癌变)进行基因芯片检测和甲基化DNA免疫沉淀测序(MeDIP-Seq),在人舌黏膜组织和人舌癌细胞系中,用实时定量逆转录聚合酶链反应(qRT-PCR)和飞行质谱检测验证转化生长因子贝塔信号蛋白1(SMAD1)的表达和启动子的甲基化。结果28周较12周和0周舌黏膜的胞嘧啶鸟嘌呤岛(CGI)甲基化水平均升高,12周时启动子甲基化水平高于0周。在0、12和28周期间,208个差异表达基因与启动子中的差异甲基化呈负相关。与正常黏膜相比,细胞系中SMAD1的mRNA上调,同时启动子甲基化水平降低。结论舌黏膜癌变中伴随DNA甲基化修饰异常,舌癌中SMAD1高表达伴启动子低甲基化。Objective This study aims to assess the role of DNA methylation changes in tongue cancer through a comprehensive analysis of global DNA methylation alterations during experimental lingual carcinogenesis.Methods C57BL/6J mice were subjected to 16-week oral administration of 4-nitroquinoline-1-oxide(4NQO,50 mg/L).Lingual mucosa samples,being representative of normal tissue(week 0)and early(week 12)and advanced(week 28)tumorigenesis,were harvested for microarray and methylated DNA immunoprecipitation sequencing(MeDIP-Seq).The mRNA and promoter methylation of transforming growth factor-beta-signaling protein 1(SMAD1)were evaluated with realtime quantitative reverse transcription polymerase chain reaction and Massarray in human lingual mucosa and tongue cancer cell lines.Results The cytosine guanine island(CGI)methylation level observed at 28 weeks surpassed that of both 12 weeks and 0 weeks.The promoter methylation level at 12 weeks exceeded that at 0 weeks.Notably,208 differentially expressed genes were negatively correlated to differential methylation in promoters among 0,12,and 28 weeks.The mRNA of SMAD1 was upregulated,concurrent with a decrease in promoter methylation levels in cell lines compared to normal mucosa.Conclusion DNA methylation changed during lingual carcinogenesis.Overexpression of SMAD1 was correlated to promoter hypomethylation in tongue cancer cell lines.

关 键 词：舌癌 发病机制 甲基化 动物模型 甲基化DNA免疫沉淀测序 差异表达基因 

分 类 号：R739.86[医药卫生—肿瘤]