基于加权基因共表达网络分析挖掘茯苓抗食管癌作用靶点

Therapeutic target exploration of Poria cocos for preventing esophageal cancer based on weighted gene co-expression network analysis

该研究利用加权基因共表达网络分析(WGCNA)探讨茯苓抗食管癌的潜在作用靶点,为食管癌的临床诊治开创新思路.首先在中药系统药理学数据库与分析平台(TCMSP)数据库中筛选出茯苓的有效活性成分及对应靶点,运用Cytoscape软件中鉴定候选药物作用靶点,接着在癌症基因组图谱(TCGA)数据库和基因表达谱差异(GEO)数据库中下载肿瘤相关基因表达数据集,通过WGCNA鉴定出与食管癌紧密相关的基因,并将鉴定出的基因与茯苓的候选药物作用靶点进行比对,确定茯苓抗食管癌的潜在作用靶点,最后采用RT-PCR的方法对筛选出核心基因在人食管癌细胞中的表达量进行检测.结果共获得34种茯苓有效活性成分,及17个对应靶点,主要候选药物作用靶点为:CHRM1、NCOA2、PGR、ADRA1B、ADH1B、PTGS2.其中ADH1B为茯苓抗食管癌的潜在作用靶点.筛选出DLGAP5、MCM2、CCNA2、CDK1、TPX2、TRIP13、KIF23、KIF2C、CENPF、CHAF1A等10个核心基因,其中CHAF1A和MCM2的表达水平对食管癌患者的预后具有影响.RT-PCR结果显示,CDK1、CHAF1A、TPIP13和MCM2基因经不同浓度的茯苓水煎液处理后,表达量与对照存在显著差异(p<0.05),表明CDK1、CHAF1ATPIP13和MCM2是茯苓水煎液治疗食管癌的主要作用靶点.

Weighted gene co-expression network analysis was used to identify the hub genes related to the occurrence and development of esophageal cancer for exploring the potential targets of anti-esophageal cancer effects of Poria cocos,and open up new avenues for the clinical diagnosis and treatment of esophageal cancer.Firstly,the effective active components of Poria cocos and their corresponding targets were screened out from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).The protein interaction network map of the targets was constructed with the STING tool,and the first 6 candidate drug action targets were identified by Cytoscape.The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)were used to explore the differentially expressed genes between normal tissues and esophageal cancer tissues.Genes related to esophageal cancer were obtained by WGCNA.The potential targets of anti-esophageal cancer effects were preliminarily determined by comparing the esophageal cancer-related genes with the first six drug targets of Poria cocos.The RT-PCR method was used to detect the expression of the selected hub genes in mouse model cancer cells.A total of 34 kinds of effective active components of Poria cocos as well as 17 corresponding targets were generated,among which the main candidate drug targets were CHRM1,NCOA2,PGR,ADRA1B,ADH1B,and PTGS2.ADH1B is demonstrated to be a potential therapeutic target for anti-esophageal cancer of Poria cocos.A total of 2,394 differentially expressed genes were screened out in the TCGA-ESCA data set,and 1,504 differentially expressed genes were screened out in the GSE22954 dataset.Hub genes DLGAP5,MCM2,CCNA2,CDK1,TPX2,TRIP13,KIF23,KIF2C,CENPF,and CHAF1A were screened out.The expression level of CHAF1A and MCM2 influences the prognosis of patients with esophageal cancer.RT-PCR results showed that the expression of CDK1,CHAF1A,TPIP13,and MCM2 genes under treatment with different concentrations of Poria cocos extracts was significantly different from the control,indicating that CDK1,CHAF1ATPIP13,and MCM2 are the main target during Poria cocos in the treatment of esophageal cancer.