许修颖,龚荣府,杨娉娉,方文
XU Xiuying,GONG Rongfu,YANG Pingping,FANG Wen

• 1:贵州医科大学医学检验学院生化教研室

摘要(Abstract)：

目的:运用生物信息学方法筛选宫颈癌(CC)枢纽基因,并挖掘CC预后分析的潜在分子标志。方法:选取基因表达数据库(GEO)中CC表达数据集,利用R语言中的软件包筛选CC组织与正常宫颈组织中存在的差异表达基因(DEGs);并进行基因本体(GO)分析、基因组百科全书(KEGG)通路分析、蛋白质相互作用(PPI)网络构建、Cytoscape软件寻找枢纽基因、The Kaplan Meier plotter网站绘制枢纽基因生存曲线图,采用肿瘤基因组图谱(TCGA)数据库的UALCAN在线数据库验证CC患者枢纽基因的表达,采用人类蛋白质图谱(HPA)数据库分析相关蛋白的免疫组织化学结果。结果:鉴定219个上调基因和182个下调基因,KEGG通路分析结果显示差异表达基因主要富集在细胞周期、DNA复制、卵母细胞减数分裂、p53信号通路及花生四烯酸代谢等通路;筛选出的枢纽基因CDK1、KIF11、CCNB1、CDC45及CXCL8在CC中表达上调(P <0. 05),并与CC患者总体生存率(OS)相关(P <0. 05),同时在TCGA数据库中枢纽基因的表达趋势与GEO数据库中相同(P <0. 01),免疫组织化学法结果显示CXCL8未被检测到,CDK1、KIF11、CCNB1及CDC45在CC组织中表达高于正常宫颈组织。结论:筛选出CDK1、KIF11、CCNB1、CDC45及CXCL8等5个CC枢纽基因,其表达与CC的预后相关。
Objective: To screen cervical cancer( CC) hub genes with bioinformatics methods and to explore potential molecular markers for prognostic analysis of CC. Methods: The CC expression data set was downloaded from the gene expression omnibus( GEO) database,by using the package in the R language to screen differentially expressed genes( DEGs) from CC and cervical normal tissues,and systematic bioinformatics analysis of DEGs was performed,including gene ontology( GO) analysis and kyoto encyclopedia of genes and genomes( KEGG) pathway analysis,protein-protein interactions( PPI) network construction. The cytoscape software was applied to search for hub genes,and the Kaplan Meier plotter website to draw hub genes survival curve graphs. In addition,the UALCAN online database based on the cancer genome atlas( TCGA) database was used to verify the expression of pivotal genes in patients with CC,and the human protein atlas( HPA) database to analyze the immunohistochemical results of related proteins. Result: 219 up-regulated genes and 182 downregulated genes were identified. KEGG pathway analysis results showed that DEGs were mainly enriched in the cell cycle,DNA replication,oocyte meiosis,p53 signaling pathway and arachidonic acid metabolism and other pathways. The selected hub genes CDK1,KIF11,CCNB1,CDC45,and CXCL8 were up-regulated in CC( P < 0. 05),and related to the overall survival rate( OS) of patients with CC( P < 0. 05). The expressional trends of the pivot genes in the TCGA database were the same as in the GEO database. Immunohistochemical results showed that CXCL8 was not detected,and the expressions of CDK1,KIF11,CCNB1,and CDC45 in CC tissues were higher than those in normal cervical tissues. Conclusion: Five hub genes of CC,namely CDK1,KIF11,CCNB1,CDC45,and CXCL8 are screened,and their expressions are related to the prognosis of CC.

关键词(KeyWords)： 宫颈肿瘤;计算生物学;预后;差异表达基因;枢纽基因;KEGG通路
cervical cancer(CC);computational biology;prognosis;differentially expressed genes(DEGs);hub genes;KEGG pathway

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81560481)

作者(Author): 许修颖,龚荣府,杨娉娉,方文
XU Xiuying,GONG Rongfu,YANG Pingping,FANG Wen

DOI: 10.19367/j.cnki.2096-8388.2020.12.004

参考文献(References)：

• [1] BRAY F,FERLAY J,SOERJOMATARAM I,et al.Global cancer statistics 2018:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185countries[J]. CA Cancer J Clin,2018,68(6):394-424.
• [2]谢珊艳，任鹏．宫颈癌发病年轻化的趋势分析与相应对策[J]．中医药管理杂志，2018,26(5):10-12.
• [3]SIEGEL R L,MILLER K D,JEMAL A. Cancer statistics,2019[J]. CA Cancer J Clin,2019,69(1):7-34.
• [4] DE MARTEL C,PLUMMER M,VIGNAT J,et al.Worldwide burden of cancer attributable to HPV by site,country and HPV type[J]. Int J Cancer,2017,141(4):664-670.
• [5] VOGELSTEIN B,PAPADOPOULOS N,VELCULESCU V E,et al. Cancer genome landscapes[J]. Science,2013,339(6127):1546-1558.
• [6]YANG W,ZHAO X,HAN Y,et al. Identification of hub genes and therapeutic drugs in esophageal squamous cell carcinoma based on integrated bioinformatics strategy[J]. Cancer Cell Int,2019,19:142.
• [7]孙东雷，赵田禾，赵曼羽，等．脂肪酸合成酶作为癌症治疗靶点的生物信息学分析[J]．现代预防医学2020,47(3):508-513,521.
• [8]石旦，徐斌，蒋敬庭．MELK m RNA表达水平在非小细胞肺癌患者中的预后价值[J]．临床检验杂志，2020,38(5):389-394.
• [9]罗何三，吴盛喜，许鸿鹞，等．基于TCGA数据乳腺癌组织UBE2T表达临床意义分析[J]．中华肿瘤防治杂志2017,24(13):901-905,911.
• [10]毛建英，袁海涵．宫颈癌进展的生物信息学研究[J]．内蒙古医科大学学报，2020,42(1):20-23.
• [11]HUANG D W,SHERMAN B T,LEMPICKI R A. Bioinformatics enrichment tools:paths toward the comprehensive functional analysis of large gene lists[J]. Nucleic Acids Res,2009,37(1):1-13.
• [12]HUANG D W,SHERMAN B T,LEMPICKI R A. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources[J]. Nat Protoc,2009,4(1):44-57.
• [13]SHANNON P,MARKIEL A,OZIER O,et al. Cytoscape:a software environment for integrated models of biomolecular interaction networks[J]. Genome Res,2003,13(11):2498-2504.
• [14]CHIN C H,CHEN S H,WU H H,et al. Cyto Hubba:identifying hub objects and sub-networks from complex interactome[J]. BMC Syst Biol,2014,8(Suppl 4):S11.
• [15]NAGY A，LANCZKY A,MENYHRT O,et al. Validation of mi RNA prognostic power in hepatocellular carcinoma using expression data of independent datasets[J]. Sci Rep,2018,8(1):9227.
• [16]CHANDRASHEKAR D S,BASHEL B,BALASUBRAMANYAS A H,et al. UALCAN:a portal for facilitating tumor subgroup gene expression and survival analyses[J].Neoplasia(New York,NY),2017,19(8):649-658.
• [17]UHLEN M,ZHANG C,LEE S,et al. A pathology atlas of the human cancer transcriptome[J]. Science,2017,357(6352):2507.
• [18]韦蒙专，陈绍俊，黄海欣．中晚期宫颈癌综合治疗模式的研究现状和进展[J]．中国肿瘤，2019,28(6):456-460.
• [19]MALUMBRES M. Cyclin-dependent kinases[J]. Genome Biology,2014,15(6):122.
• [20]KISHIMOTO T. MPF-based meiotic cell cycle control:half a century of lessons from starfish oocytes[J]. Proc Jpn Acad Ser B Phys Biol Sci,2018,94(4):180-203.
• [21]LI M,HE F,ZHANG Z,et al. CDK1 serves as a potential prognostic biomarker and target for lung cancer[J].The Journal of international medical research,2020,48(2):030006051989750.
• [22]LUO Y,WU Y,PENG Y,et al. Systematic analysis to identify a key role of CDK1 in mediating gene interaction networks in cervical cancer development[J]. Irish Journal of Medical Science,2016,185(1):231-239.
• [23]DAIGO K,TAKANO A,THANG P M,et al. Characterization of KIF11 as a novel prognostic biomarker and therapeutic target for oral cancer[J]. Int J Oncol,2018,52(1):155-165.
• [24]LIU L,LIU X,MARE M,et al. Overexpression of Eg5correlates with high grade astrocytic neoplasm[J]. Journal of Neuro-Oncology,2016,126(1):77-80.
• [25]WANG F,CHEN X,YU X,et al. Degradation of CCNB1 mediated by APC11 through UBA52 ubiquitination promotes cell cycle progression and proliferation of nonsmall cell lung cancer cells[J]. American Journal of Translational Research,2019,11(11):7166-7185.
• [26]贾未玲，于淼．Cyclin B1与肿瘤细胞周期调控[J]．哈尔滨商业大学学报(自然科学版)，2016,32(6):659-662.
• [27]杨生辉，黄琰菁，邱纯，等．基于数据库探讨细胞周期蛋白B1(CCNB1)表达对胃癌预后的影响[J]．中国数字医学，2020,15(1):37-40.
• [28]KURNIAWAN F,SHI K,KURAHASHI K,et al. Crystal structure of entamoeba histolytica Cdc45 suggests a conformational switch that may regulate dna replication[J]. i Science,2018:102-109.
• [29]PETOJEVIC T,PESAVENTO J J,COSTA A,et al.Cdc45(cell division cycle protein 45)guards the gate of the eukaryote replisome helicase stabilizing leading strand engagement[J]. Proceedings of the National Academy of ences of the United States of America,2015,112(3):249-258.
• [30]MOYER S E,LEWIS P W,BOTCHAN M R. Isolation of the Cdc45/Mcm2-7/GINS(CMG)complex,a candidate for the eukaryotic DNA replication fork helicase[J]. Proceedings of the National Academy of Sciences of the United States of America,2006,103(27):10236-10241.
• [31]BAGGIOLINI M. CXCL8-The First Chemokine[J].Front Immunol,2015,6:285.
• [32]LIU Q,LI A,TIAN Y,et al. The CXCL8-CXCR1/2pathways in cancer[J]. Cytokine Growth Factor Rev,2016,31:61-71.
• [33]YAN R,SHUAI H,LUO X,et al. The clinical and prognostic value of CXCL8 in cervical carcinoma patients:immunohistochemical analysis[J]. Bioscience Reports,2017,37(5):BSR20171021.

文章评论(Comment)：