CT表现为结节的肺腺癌患者的组织甲基化特征分析  

Feature analysis of the tissue methylation profile in adenocarcinoma patients with pulmonary nodules on CT scan

作　　者：郭巧梅 乔理华 王琳[1] 王薛庆 吴飞 梁小卉 孙玉腾 娄加陶[1] Guo Qiaomei;Qiao Lihua;Wang Lin;Wang Xueqing;Wu Fei;Liang Xiaohui;Sun Yuteng;Lou Jiatao(Department of Laboratory Medicine,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai200080,China;School of Laboratory Medicine,Bengbu Medical University,Bengbu233030,China)

机构地区：[1]上海市第一人民医院,上海交通大学医学院附属第一人民医院检验医学中心,上海200080 [2]蚌埠医科大学检验医学院,蚌埠233030

出　　处：《中华检验医学杂志》2024年第11期1277-1285,共9页Chinese Journal of Laboratory Medicine

基　　金：上海市卫健委协同创新集群项目(2019CXJQ03);上海申康医院发展中心研究型医师创新转化能力培训项目(SHDC2022CRD020);国家自然科学基金(82002224);上海市第一人民医院特色研究项目(CTCCR-2021B06)。

摘　　要：目的分析CT表现为结节的肺腺癌患者的组织甲基化特征。方法回顾性研究。收集2022年6月1日至2024年1月20日于上海市第一人民医院确诊为肺腺癌的70例肺结节患者。使用随机数字表法将其分为2组,发现组40例和验证组30例。发现组中组织样本采用简并亚硫酸氢盐测序技术(RRBS)检测分析,比较癌组织和配对癌旁组织的平均甲基化水平;筛选差异性甲基化区域(DMR),分析其在不同基因功能元件上的分布并绘制聚类热图;进一步对DMR进行GO和KEGG功能富集分析;随后对获得的潜在肺腺癌相关甲基化基因通过TCGA肺腺癌队列和靶向亚硫酸氢盐测序(TBS)在验证组中进行验证。组间甲基化水平的比较采用t检验或非参数检验,率和构成比采用χ^(2)检验或者Fisher精确检验。结果发现组中,癌组织平均甲基化水平低于癌旁组织[(42.369±4.627)比(44.370±4.046),t=-2.059,P=0.043];共筛选出37995个DMR,包括16889个高甲基化区域和21106个低甲基化区域,主要分布在启动子区(48.917%)、内含子区(36.457%)和外显子区(10.812%);DMR聚类热图显示癌组织和癌旁组织分别形成两个聚类。GO分析显示DMR相关基因主要定位在细胞膜和核染色质上,参与RNA聚合酶Ⅱ相关的转录及调控过程。KEGG通路富集分析显示DMR相关基因主要参与神经活性配体-受体相互作用通路、癌症通路、钙信号通路、cAMP信号通路和MAPK信号传导通路。TCGA队列验证出了11个潜在的特征性DMR。在验证组中采用TBS证实MIR10B、DMRTA2、HOPX、TFAP2B及MARCH11相关的DMR在癌组织中甲基化水平高于癌旁组织[11.200(4.305,27.088)比2.650(1.298,4.645),Z=-4.539,P<0.05;18.610(13.600,33.025)比8.675(5.488,13.085),Z=-4.554,P<0.05;17.600(2.183,76.015)比1.085(0.898,1.835),Z=-5.131,P<0.05;5.250(3.220,7.693)比3.495(2.165,4.383),Z=-2.861,P<0.05;11.515(7.525,21.033)比7.830(5.518,11.488),Z=-2.440,P<0.05],差异均有统计学意义。结论肺腺癌组织与�Objective To investigate the tissue methylation features of adenocarcinoma patients presenting as pulmonary nodules on CT scans.Methods A retrospective analysis was conducted on 70 adenocarcinoma patients with pulmonary nodules diagnosed at the Shanghai General Hospital from June 1,2022 to January 20,2024.Participants were assigned to two groups using the random number table,with 40 in the discovery group and 30 in the validation group.In the discovery group,tissue samples were analyzed using reduced representation bisulfite sequencing(RRBS)technology to compare the average methylation levels between cancer tissues and paired adjacent non-cancerous tissues.Differentially methylated regions(DMRs)were screened for analysis of their distribution across various genomic functional elements,and hierarchical clustering was plotted.GO and KEGG pathway enrichment analyses were further conducted on the DMRs.Subsequently,candidate DMRs associated with lung adenocarcinoma were validated using TCGA lung adenocarcinoma cohort and targeted bisulfite sequencing technology in the validation group.The comparison of methylation levels between groups was conducted using t-tests or non-parametric tests,while rates and composition ratios were analyzed using chi-square tests or Fisher′s exact test.Results In discovery cohort,the average methylation level in cancer tissues was lower compared to adjacent normal tissues[(42.369±4.627)vs(44.370±4.046),t=‒2.059,P=0.043].A total of 37995 DMRs were identified,including 16889 upregulated regions and 21106 downregulated regions,predominantly locating in promoter regions(48.917%),introns(36.457%),and exons(10.812%).The DMR clustering heatmap revealed two distinct clusters corresponding to cancer tissues and adjacent non-cancerous tissues.GO analysis showed that DMRs associated genes were mainly located in the cell membrane and nuclear chromatin,and were primarily involved in RNA polymeraseⅡ-related transcription and regulation.KEGG pathway enrichment analysis indicated that DMRs associat

关 键 词：结节病 肺 腺癌 DNA甲基化 

分 类 号：R734.2[医药卫生—肿瘤] R730.44[医药卫生—临床医学]