乳腺癌中关键基因及靶向治疗化合物的筛选  

Screening of key genes and targeted therapeutic compounds in breast cancer

作　　者：刘敏 黄健 晏娇艳 杨烨 袁艳 何芸 宋孝晗 莫非 罗昭逊[3] 张姝 LIU Min;HUANG Jian;YAN Jiaoyan;YANG Ye;YUAN Yan;HE Yun;SONG Xiaohan;MO Fei;LUO Zhaoxun;ZHANG Shu(Department of Basic Clinical Laboratory and Hematology,School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,Guizhou,China;Center for Clinical Laboratories,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;School of Pediatrics,Guizhou Medical University,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学医学检验学院临床检验基础与血液学教研室,贵州贵阳550004 [2]贵州医科大学附属医院临床检验中心,贵州贵阳550004 [3]贵州医科大学儿科学院,贵州贵阳550004

出　　处：《贵州医科大学学报》2021年第8期876-885,共10页Journal of Guizhou Medical University

基　　金：国家自然科学基金(81860723,81960589);中国博士后科学基金(2018M643862);贵州省卫生计生委科学技术基金(gzwjkj2018-1-073);中医药、民族医药科学技术基金(QZYY-2018-019);贵州省科技计划项目[黔科合支撑(2021)一般097]。

摘　　要：目的筛选并验证乳腺癌(BC)中潜在的关键基因(Hub基因),探讨其在BC中的发生机制并寻找以Hub基因为靶点治疗BC的潜在化合物。方法使用GEO2R在线工具从基因表达数据库(GEO)中获取BC中的差异表达基因(DEGs)后,利用基因表达谱数据动态分析(GEPIA)数据库和人类蛋白质图谱(HPA)数据库分别从mRNA水平和蛋白质层面进一步筛选潜在的Hub基因;采用逆转录-实时荧光定量聚合酶链式反应(qRT-PCR)、蛋白印迹(Western blot)和免疫组织化学染色法从mRNA水平和蛋白质层面对Hub基因在BC组与癌旁组的表达进行验证,绘制Hub基因的生存曲线,分析DEGs涉及的相关信号通路和调控机制,从Connectivity map(Cmap)数据库中筛选具有抗BC活性的候选化合物。结果qRT-PCR、Western blot及免疫组织化学表明11个Hub基因及编码的蛋白在BC组中表达上调,功能分析表明,MELK、CDK1、HMMR和AURKA 4个基因与细胞周期调控相关,GINS2、RRM2和CDC45基因与DNA复制或修复相关,SMC4、PRC1和TOP2A与细胞分裂相关;Cmap数据库分析共鉴定出6个具有抗BC活性的候选化合物。结论筛选出11个Hub基因,其表达与BC患者预后相关。Objective To screen and verify the potential key genes(Hub genes)in breast cancer(BC),explore their mechanism in BC,and find potential compounds targeting Hub genes for the BC therapy.Methods GEO2R online tool was used to obtain the differentially expressed genes(DEGs)in BC from the gene expression database(GEO).The gene expression profile interactive analysis(GEPIA)database and the human protein atlas(HPA)database were used to obtain mRNA levels and protein levels of genes of interest to screen Hub genes.Real-time fluorescent quantitative polymerase chain reaction(qRT-PCR),Western blot and immunohistochemical(IHC)staining were used to detect the Hub genes in the BC group and adjacent cancers at mRNA level and the protein level.Survival curves of the Hub genes was drawn.DEGs-involved signal pathways and regulatory mechanisms were analyzed.Candidate compounds with anti-BC activity were screened from the Connectivity map(Cmap)database.Results qRT-PCR,Western blot,and IHC staining showed that 11 Hub genes and their encoded proteins were up-regulated in the BC group.Functional analysis showed that MELK,CDK1,HMMR,and AURKA were related to cell cycle regulation,and GINS2,RRM2,CDC45 were related to DNA replication or repair,and SMC4,PRC1,and TOP2A were related to cell division.Cmap database analysis identified 6 candidate compounds with anti-BC activity.Conclusion The expression levels of identified 11 Hub genes are associated with the prognosis of BC patients.

关 键 词：基因 乳腺肿瘤 化合物 生物信息学 生存曲线 

分 类 号：R737.9[医药卫生—肿瘤] Q811.4[医药卫生—临床医学]