基于基因表达数据库的急性髓系白血病细胞焦亡预后分析及GZMB调节机制研究  

Prognostic analysis of pyroptosis and molecular mechanism of GZMB in acute myeloid leukemia based on TCGA and GTEx database

作　　者：党庆秀 李洋 郭嫣婷 沈洋灵 顾伟英[1] DANG Qingxiu;LI Yang;GUO Yanting;SHEN Yangling;GU Weiying(Department of Hematology,the Third Affiliated Hospital of Soochow University,the First People's Hospital of Changzhou,Jiangsu Changzhou 213003,China;The School of Medicine,Jiangsu University,Jiangsu Zhenjiang 212031,China)

机构地区：[1]苏州大学附属第三医院,常州市第一人民医院血液科,江苏常州213003 [2]江苏大学医学院,江苏镇江212031

出　　处：《现代肿瘤医学》2023年第18期3376-3383,共8页Journal of Modern Oncology

基　　金：江苏省常州市十四五卫生领军人才工程资助项目(编号:KY20221336);江苏省卫生健康委员会重点项目(编号:ZD2021043)。

摘　　要：目的:通过生物信息分析与实验相结合探讨急性髓系白血病(acute myeloid leukemia,AML)细胞焦亡相关基因在AML患者的预后价值,并验证AML治疗的新靶点。方法:在基因表达谱数据库(TCGA、GTEx)中选取AML细胞焦亡相关基因表达数据,采用R语言筛选AML患者和健康对照组中细胞焦亡差异表达基因。对差异的基因进行单因素Cox及Lasso回归分析并构建预后模型。应用单因素和多因素Cox回归分析风险模型、临床特征与AML患者预后的相关性。Kaplan-Meier法分析预后模型中GZMB表达量与患者生存率的关系,将AML患者分成GZMB高表达组和低表达组并进行基因集富集分析(GSEA)。并采用Western blot验证AML中GZMB/Caspase-3/GSDME介导的细胞焦亡。结果:45个焦亡相关基因在AML患者组和正常组中存在差异表达(P<0.05)。其中7个基因(BAK1、CASP3、GZMB、GPX4、NLRP2、GSDMB和GSDMA)被纳入风险模型构建。年龄、FAB分型、风险评分与AML患者预后相关(P<0.05),且风险评分是AML的独立危险因素(P<0.05)。GZMB高表达组的总生存率显著低于低表达组,差异有统计学意义(P<0.05)。GSEA富集分析显示GZMB主要参与了细胞因子信号通路、NK细胞、B细胞、T细胞免疫调节等信号途径(P<0.05,FDR≤25%)。Western blot实验结果初步证实AML中存在GZMB/Caspase-3/GSDME介导的细胞焦亡调节途径。结论:通过生物信息学方法,构建了7个细胞焦亡相关基因的AML预后模型,GZMB高表达与AML发生发展及不良预后相关,并参与GZMB/Caspase-3/GSDME途径介导的细胞焦亡。Objective:Bioinformatics analysis was used to screen differentially expressed genes(DEGs)in acute myeloid leukemia(AML)and further validation and analysis of their prognostic value were performed,in order to identify and validate novel targets for AML therapy.Methods:The expression data and clinical characteristics of AML and normal samples were downloaded from TCGA and GTEX database.R software was used to screen the DEGs in tumor tissues and healthy controls.The DEGs were identified by Cox regression analysis and prognostic analysis was carried out by Lasso analysis.Univariate and multivariate Cox regression were applied to analyze the correlation between risk model,clinical traits and prognosis of AML patients.Kaplan-Meier method was used to analyze the relationship between GZMB expression and survival rate in the prognosis model.AML patients were divided into high GZMB expression group and low GZMB expression group,and gene set enrichment analysis(GSEA)was performed.Western blot and ELISA were used to verify GZMB/Caspase-3/GSDME-mediated cell scorching in AML.Results:Forty-five DEGs were analyzed for prognosis,and 7 genes(BAK1,CASP3,GZMB,GPX4,NLRP2,GSDMB and GSDMA)were included in the risk model construction.Age,FAB subtypes,and risk score were significantly associated with the prognosis of AML patients.The overall survival rate of the group with high expression of GZMB was lower than that of the group with low(P<0.05).GSEA enrichment analysis showed that GZMB was mainly involved in cytokine signaling pathway,natural killer cell,B cell and T cell immune regulation signaling pathway(P<0.05,FDR≤25%).Western blot results showed the participation of pyroptosis mediated by GZMB/Caspase-3/GSDME in AML.Conclusion:The prognostic model of AML constructed by bioinformatic analyses with seven cell scorch-related genes may provide reference for the individualized treatment and evaluation of AML patients.The GZMB/Caspase-3/GSDME-mediated pyroptosis is involved in AML.

关 键 词：急性髓系白血病 TCGA GTEx 细胞焦亡 GZMB 

分 类 号：R733.71[医药卫生—肿瘤]