天津医药

天津医药 ›› 2018, Vol. 46 ›› Issue (6): 610-614.doi: 10.11958/20171427

• 细胞与分子生物学 • 上一篇    下一篇

基于基因芯片探索鼻咽癌的分子靶标

刘玉智，门剑龙，李杨   

  1. 1天津医科大学总医院耳鼻喉科（邮编300052），2精准医学中心
  • 收稿日期:2017-12-11 修回日期:2018-03-12 出版日期:2018-06-15 发布日期:2018-07-05
  • 通讯作者: 李杨 E-mail:yangli.tijmu@foxmail.com

Identify the molecular target of nasopharyngeal carcinoma by bioinformatics analysis

LIU Yu-zhi, MEN Jian-long, LI Yang   

  1. 1 Department of Otolaryngology, 2 Precision Medicine Center, Tianjin Medical University General Hospital, Tianjin 300052, China
  • Received:2017-12-11 Revised:2018-03-12 Published:2018-06-15 Online:2018-07-05

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摘要： 目的 在分子水平探索鼻咽癌的发病机制，筛选鼻咽癌诊断或治疗的潜在分子靶标。方法 在GEO公共数据库中下载编号为GSE12452和GSE13597的基因芯片数据（包含鼻咽癌和对照鼻炎黏膜组织数据），利用R编程语言的相关工具包对原始芯片数据进行预处理和差异表达基因的筛选。利用DAVID数据库对差异表达基因进行基因GO功能分析和KEGG信号通路分析。取我院鼻咽癌石蜡标本和对照鼻炎黏膜组织新鲜标本各5例，利用realtime PCR和Western blot分别检测18个细胞周期相关基因和4种蛋白在两组织中的表达，进一步验证基因芯片的结果。结果 生物信息学分析得到了260个差异表达基因，涉及16个GO条目和4条信号通路。18个细胞周期相关的基因中，real-time PCR和Western blot进一步确定鼻咽癌组织中有12个基因在mRNA水平表达上调，4个基因在蛋白水平表达上调。结论 通过对两套鼻咽癌表达谱芯片数据的综合生物信息学分析与分子生物学验证，发现CDC6、CDK1、MCM2和CCNB1这4个可能是鼻咽癌恶性进展相关的关键基因，可作为潜在靶点作进一步的功能研究。

关键词: 鼻咽肿瘤, 细胞周期, 寡核苷酸序列分析, 计算生物学, 鼻咽癌, GEO数据库, DAVID数据库

Abstract: Objective To study the pathogenesis of nasopharyngeal carcinoma and identify potential biomarkers or therapeutic targets. Methods Microarray data (GSE12452 and GSE13597) were downloaded from Gene Expression Omnibus. Processing of original microarray data and screening of differentially expressed genes were performed through bioinformatics analysis. Then, GO and KEGG pathway enrichment analysis was performed for these genes using DAVID database. Real time-PCR and Western blot assay were used to detect the expression levels of the identified genes. Results A total of 260 overlap DEGs were obtained including 16 GO entries and 4 signal pathways. Eighteen potential therapeutic targets that relative to cell cycle were identified by gene enrichment analysis. Expression levels of 12 selected genes were confirmed by real-time PCR. Finally, 4 selected genes were confirmed by Western blot assay. Conclusion By bioinformatics analysis of two sets of microarray data and molecular biology research, four genes were found including CDC6，CDK1，MCM2 and CCNB1, which might be potential key genes that can be developed for therapy targets of NPC in the future

Key words: nasopharyngeal neoplasms, cell cycle, oligonucleotide array sequence analysis, computational biology, nasopharyngeal carcinoma, Gene Expression Omnibus database, DAVID database