早期甲状腺乳头状癌颈部淋巴结转移相关分子标志物的转录组测序分析  

Transcriptome sequencing analysis of molecular markers related to cervical lymph node metastasis in early stage of papillary thyroid carcinoma

作　　者：杨帆[1] 廉猛[2] 陈佳铭 王茹[2] 张嘉敏 沈茜茜 冯凌[2] 孟令照[1] 房居高[2] YANG Fan;LIAN Meng;CHEN Jiaming;WANG Ru;ZHANG Jiamin;SHEN Xixi;FENG Ling;MENG Lingzhao;FANG Jugao(Department of Otolaryngology Head and Neck Surgery,Beijing Anzhen Hospital,Capital Medical University,Beijing,100029,China;Department of Otolaryngology Head and Neck Surgery,Beijing Tongren Hospital,Capital Medical University,Key Laboratory of Otolaryngology Head and Neck Surgery(Capital Medical University),Ministry of Education,Beijing,100730,China)

机构地区：[1]首都医科大学附属北京安贞医院耳鼻咽喉头颈外科,北京100029 [2]首都医科大学附属北京同仁医院耳鼻咽喉头颈外科,耳鼻咽喉头颈科学教育部重点实验室(首都医科大学),北京100730

出　　处：《中国耳鼻咽喉头颈外科》2020年第12期673-678,共6页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：北京市医院管理局“登峰”人才培养计划(DFL20180202);北京市中医局科技发展资金项目(QN2018-32);首都卫生发展科研专项自主创新项目(2018-2-2054);吴阶平基金(MTEK-TB20170627)。

摘　　要：目的使用转录组测序技术筛选早期甲状腺乳头状癌(papillary thyroid carcinoma,PTC)与淋巴结转移相关的差异表达基因,使用生物信息学技术进一步探讨和验证具有关键调控预测作用的功能性靶基因。方法应用转录组测序技术对7例出现颈部淋巴结转移和9例无颈部淋巴结转移的早期(T1级)PTC肿瘤样本进行差异表达基因筛选,并进一步对差异表达基因进行生物信息学验证,寻找早期PTC淋巴结转移潜在分子标志物。结果转录组测序共筛选出1805个差异表达基因,其中上调1049个,下调756个。经过GO数据库分析,差异基因主要聚类在细胞代谢、黏附、分裂增殖等集合中(P<0.05);KEGG数据库分析发现差异基因显著富集于癌症途径通路、黏附作用通路、PI3K-Akt信号通路和氧化磷酸化等通路中(P<0.05)。分别构建蛋白质-蛋白质互作网络和随机森林后,发现差异表达基因FN1、ADCY7、LDLR、NOS1和GNA13既处于互作网络核心区域同时在随机森林分类中对淋巴结转移具有显著特征选择价值。TCGA数据库验证发现差异基因FN1在伴淋巴结转移早期PTC中显著低表达(P<0.001),LDLR(P<0.001)、NOS1(P<0.05)、GNA13(P<0.05)在伴淋巴结转移早期PTC中显著高表达,且通过绘制ROC曲线和计算曲线下面积发现这4个基因对早期PTC淋巴结转移有较高的预测价值。结论伴/不伴有淋巴结转移早期PTC在整体基因表达上存在较明显的差异,其中差异表达基因FN1、LDLR、NOS1和GNA13具有作为早期PTC淋巴结转移相关分子标志物的潜在价值。OBJECTIVE Transcriptome sequencing technique was used to screen differentially expressed genes associated with lymph node metastasis in early papillary thyroid carcinoma(PTC),and bioinformatics techniques were used to further explore and verify the functional target genes that play a key role in regulation and prediction.METHODS A total of 16 early-stage PTC tissue samples,of which 7 with cervical lymph node metastasis and 9 without cervical lymph node metastasis,were included and divided into two group according to the existence of cervical lymph node metastasis.Gene expression were detected by RNA-sequencing and were validated by bioinformatic analysis.RESULTS A total of 1805 differentially expressed genes related to cervical lymph node metastasis were detected,1049 genes were up-regulated and 756 genes were down-regulated.Analyzed by GO database,the functional sets were consisted of cell metabolism,cell adhesion and cell proliferation(P<0.05).Functional pathways including pathways in cancer,focal adhesion,PI3K-Akt signaling pathway and oxidative phosphorylation pathway were found enriched in differentially expressed genes based on the enrichment analysis via KEGG database(P<0.05).After construction of protein-protein interaction(PPI)network and random forest classifier,differentially expressed gene FN1,LDLR,NOS1 and GNA13 were identified as with predicting potentials of cervical lymph node metastasis in early-stage PTC.Further validation confirmed our findings by comparing the expression level and drawing the ROC curves of FN1,LDLR,NOS1 and GNA13 based on the TCGA database.CONCLUSION There were significantly statistical differences in the overall pattern of gene expression between early-stage PTCs with/without cervical lymph node metastasis.As potential biomarkers,FN1,LDLR,NOS1 and GNA13 may play important roles in the predicting cervical lymph node metastasis of early-stage PTC.

关 键 词：甲状腺肿瘤 癌 乳头状 聚类分析 淋巴结转移 转录组测序 生物信息学分析 

分 类 号：R736.1[医药卫生—肿瘤]