SHOX2和RASSF1A基因启动子区甲基化检测在早期肺腺癌筛查和诊断中的应用价值  

Application value of SHOX2 and RASSF1A gene promoter region methylation detection for screening and diagnosis of early-stage lung adenocarcinoma

作　　者：丁杰[1] 葛美玲[1] 胡月[1] 刘艳红[1] 王晓巍 高玒[1] Ding Jie;Ge Meiling;Hu Yue;Liu Yanhong;Wang Xiaowei;Gao Hong(Department of Biobank,Nanjing Drum Tower Hospital,the Affiliated Hospital of Nanjing University Medical School,Nanjing 210008,China)

机构地区：[1]南京大学医学院附属鼓楼医院生物样本库,南京210008

出　　处：《肿瘤研究与临床》2023年第3期185-192,共8页Cancer Research and Clinic

基　　金：江苏省重大疾病生物资源样本库项目(BM2015004);江苏省重大疾病生物资源样本库开放课题(TC2021B012、TC2021B020、TC2021B021、TC2021B022)。

摘　　要：目的探讨SHOX2和RASSF1A基因启动子区甲基化检测对早期肺腺癌筛查及诊断的价值。方法下载癌症基因组图谱(TCGA)数据库中471例肺腺癌患者的mRNA测序数据及相应的413例甲基化数据分别计算两基因启动子区甲基化水平,分析正常肺组织与肿瘤组织的甲基化水平差异。回顾性分析2018年1月至2019年1月南京大学医学院附属鼓楼医院接受手术的54例早期肺腺癌及31例肺良性肿瘤患者临床资料,检测肿瘤组织及正常肺组织(距肿瘤>5 cm)(称为:临床样本)SHOX2及RASSF1A甲基化状态,以任一基因启动子区甲基化阳性为两基因联合甲基化阳性。以病理诊断为金标准,绘制受试者工作特征(ROC)曲线,分析基因甲基化阳性诊断早期肺腺癌的效能。筛选出石蜡样本中肿瘤细胞比例>80%的患者,对其肿瘤组织及正常肺组织进行mRNA高通量测序。分析两基因甲基化阳性与患者临床病理特征的关系,采用Spearman法分析临床样本和TCGA数据库样本中两基因启动子区甲基化水平与mRNA表达水平的相关性。采用基因集变异分析(GSVA)法分析两基因甲基化阳性临床肺腺癌样本和对应的甲基化阴性肺腺癌样本间京都基因与基因组百科全书(KEGG)富集通路的差异。结果在TCGA数据库中,SHOX2启动子区甲基化岛包含6个被测序的甲基化位点,其中位点cg04532033、cg01557547甲基化水平在肺腺癌组织中均高于正常肺组织(均P<0.05);RASSF1A基因启动子区甲基化岛包含11个被测序的甲基化位点,其中6个位点的甲基化水平在肺腺癌组织中均高于正常肺组织(均P<0.05);与正常肺组织比较,SHOX2启动子区甲基化水平在Ⅰ、Ⅱ期肺腺癌组织中升高(均P<0.05);RASSF1A启动子区甲基化在肺腺癌各期中均处于较高水平(均P<0.001)。在临床54例早期肺腺癌患者中,癌组织SHOX2启动子区甲基化阳性28例,RASSF1A启动子区甲基化阳性21例,两基因联合甲基化阳性40例;31例Objective To investigate the value of SHOX2 and RASSF1A gene promoter region methylation detection for screening and diagnosis of early-stage lung adenocarcinoma.Methods The mRNA sequencing data of 471 lung adenocarcinoma patients and corresponding methylation data of 413 cases were downloaded from The Cancer Genome Atlas(TCGA)database,the methylation levels of SHOX2 and RASSF1A gene promoter regions were calculated,and the difference in methy lation level between normal lung tissues and tumor tissues was analyzed.The clinical data of 54 patients with early-stage lung adenocarcinoma and 31 patients with benign lung tumors who underwent surgery at Drum Tower Hospital Affiliated to Nanjing University Medical School from January 2018 to January 2019 were retrospectively analyzed.The methylation status of SHOX2 and RASSF1A in tumor tissues and normal lung tissues(>5 cm from the edge of the tumor foci)(called clinical samples)was detect,and a positive methylation in the promoter region of either gene was considered as a combination of two genes methylation positivity.Using pathological diagnosis as the gold standard,the efficacy of gene methylation positivity in diagnosing early-stage lung adenocarcinoma was analyzed by receiver operating characteristic(ROC)curve.Patients with>80%of tumor cells in paraffin samples were screened,and mRNA high-throughput sequencing was performed in their tumor tissues and normal lung tissues.The relationship between positive methylation of the two genes and clinicopathological features was analyzed,and the correlation between the promoter region methylation level of the two genes and mRNA expression levels in clinical samples and TCGA database samples was analyzed by Spearman method.Gene set variance analysis(GSVA)method was used to analyze the differences in Kyoto Encyclopedia of Genes and Genomes enrichment pathways between two-gene methylation-positive clinical lung adenocarcinoma samples and corresponding methylation-negative lung adenocarcinoma.Results In TCGA database,the SHOX2 pr

关 键 词：肺肿瘤 腺癌 DNA甲基化 SHOX2 RASSF1A 

分 类 号：R734.2[医药卫生—肿瘤]