1、山东医药2023 年第 63 卷第 3 期弥漫大B细胞淋巴瘤预后相关基因与患者预后及肿瘤浸润性免疫细胞比例的相关性郭佳丽1，潘晨华2，李依3，刘晓倩4，蒋慧4，常伟41 武汉科技大学医学院武汉科技大学附属华润武钢总医院血液内科，武汉 430080；2 武汉科技大学附属普仁医院分子医学实验室；3 赤峰市医院血液内科；4 武汉科技大学附属普仁医院血液科摘要：目的筛选出弥漫大B细胞淋巴瘤（DLBCL）预后相关基因，明确预后相关基因表达水平与DLBCL患者预后及肿瘤浸润性免疫细胞（TICs）比例的关系，为DLBCL治疗提供新的靶点。方法从GEO中获取GSE56315、GSE12453和GSE87371
2、芯片数据，用R的“limma”包筛选出DLBCL预后相关基因。将GSE87371数据集中的预后相关基因根据表达量中位值分为高、低表达组，从R语言的“survival”包中提取高、低表达组5年OS率及95%CI，然后利用COX风险回归模型分析预后相关基因与预后的相关性，使用R语言的“for”循环分析预后不良相关基因与DLBCL患者年龄、性别、临床分期等临床参数的关系。通过GSEA富集分析DLBCL预后相关基因在DLBCL中的相关信号通路。最后利用CIBERSORT算法计算DLBCL组织中 TICs的比例，并分析预后相关基因表达水平与TICs比例的关系。结果对GSE12353及GSE56315进行
3、合并后的MERGE基因集进行筛选后获得DLBCL组织与正常组织的差异表达基因445个（上调基因301个，下调基因143个），从GSE87371数据集获得DLBCL预后相关基因57个（负相关17个，正相关40个），将上调的301个差异表达基因与负相关的17个基因取交集，获得膜连蛋白1（ANXA1），即为DLBCL预后相关基因。ANXA1高表达组 5年 OS率 3.56%，95%CI 1.17%10.7%，低表达组 5年 OS率 17.49%，95%CI 为11.8%29.9%，ANXA1高表达组DLBCL患者5年OS率低于低表达组（P0.001）；ANXA1为 DLBCL独立预后影响因子（P0.
4、05）。ANXA1在趋化因子信号通路、MAPK信号通路、ECM受体相互作用、JAK-STAT信号通路等通路上富集。DLBCL组织中T细胞和巨噬细胞比例较大，分别为49%、31%；ANXA1表达水平与巨噬细胞M2、浆细胞、活性CD4+记忆T细胞、CD8+T细胞、T细胞比例正相关（P均0.001）。结论DLBCL预后相关基因为ANXA1，ANXA1表达水平高与DLBCL患者预后不良及肿瘤免疫浸润细胞比例相关，富集的信号通路主要有趋化因子信号通路等肿瘤相关信号通路。关键词：弥漫大B细胞淋巴瘤；ANXA1；肿瘤免疫浸润细胞doi：10.3969/j.issn.1002-266X.2023.03.007
5、 中图分类号：R551.2 文献标志码：A 文章编号：1002-266X（2023）03-0033-04Correlation of independent prognosis-related genes in diffuse large B-cell lymphoma with patients prognosis and proportion of tumour-infiltrating immune cellsGUO Jiali1，PAN Chenhua，LI Yi，LIU Xiaoqian，JIANG Hui，CHANG Wei1 School of Medicine，Wuhan Un
6、iversity of Science and Technology，Wuhan 430080，ChinaAbstract：Objective To screen out the prognosis-related genes in diffuse large B-cell lymphoma（DLBCL）and to confirm the relationship between the expression of prognosis-related genes and patients survival and the proportion of tumour-infiltrating i
7、mmune cells（TICs）to provide new targets for the treatment and prognosis of DLBCL.Methods GSE56315，GSE12453 and GSE87371 microarray data were obtained from GEO，and DLBCL prognosis-associated genes were screened using the R limma package.Prognosis-related genes were analyzed for survival，independent p
8、rognosis，and correlation between prognosis-related gene expression levels and clinical parameters.The target genes in the GSE87371 datasets were categorised into high and low expression groups according to their median expression values，and the 5-year OS rates and 95%CIs of the high and low expressi
9、on groups were extracted from the survival package in R.基金项目：湖北省卫生健康委联合基金项目（WJ2018H0113）；武汉市卫生计生委医学科研项目（WX21C23）；武汉市卫生计生委医学科研项目（WX18C32）。第一作者简介：郭佳丽（1997-），女，在读硕士研究生，主要研究方向为血液肿瘤发病机制的研究。E-mail：通信作者简介：常伟（1972-），男，副主任医师，主要研究方向为血液系统疾病及血液肿瘤的诊治及相关基础研究。E-mail：开放科学（资源服务）标识码（OSID）33山东医药2023 年第 63 卷第 3 期The co
10、rrelation between the target genes and prognosis was then analyzed using COX risk regression models.Finally，for loop was used to analyse the relationship between the target gene and clinical parameters such as age，gender and clinical stage of DLBCL patients.The functional pathways of DLBCL prognosis
11、-related genes in DLBCL were analyzed by GSEA enrichment.Finally，the CIBERSORT algorithm was used to calculate the proportion of TICs in DLBCL tissues and to analyse the relationship between the expression levels of prognosis-related genes and the proportion of TICs.Results The merged MERGE gene set
12、 of GSE12353 and GSE56315 was screened.We obtained 445（301 up-regulated genes and 143 down-regulated genes）differentially expressed genes；57 prognosis-related genes（17 negatively and 40 positively correlated）were obtained from the GSE87371 dataset，and the 301 differentially up-regulated genes and th
13、e 17 negatively correlated genes were intersected to obtain membrane linked protein 1（ANXA1），which was a DLBCL prognosis-associated gene.Survival analysis suggested a 5-year OS rate of 3.56%（95%CI 1.17%-10.7%）in the high ANXA1 expression group and 17.49%（95%CI 11.8%-29.9%）in the low ANXA1 expression
14、 group，suggesting a poor prognosis for DLBCL patients in the high ANXA1 expression group（P0.001）.ANXA1 was an independent prognostic factor（P0.05）.GSEA functional enrichment analysis showed that the ANXA1 high expression group was enriched in chemokine signalling pathway，MAPK signalling pathway，ECM
15、receptor interaction and JAK-STAT signalling pathway.Immuno-infiltration analysis showed that T cells（49%）and macrophages（31%）were more predominant in DLBCL tissues，and ANXA1 expression was positively correlated with macrophage M2，plasma cells，activated CD4+memory T cells，CD8+T cells，and T cells.Con
16、clusion ANXA1 is associated with the prognosis of DLBCL patients，and its high expression is related to the poor prognosis and the proportion of TICs；the enrichment signal pathway mainly includes tumor-related signaling pathways such as the chemokines signaling pathway.Key words：diffuse large B-cell lymphoma；ANXA1；tumour-infiltrating immune cells弥漫大B细胞淋巴瘤（DLBCL）是成人中最常见的淋巴恶性肿瘤，具有高度的侵袭性及异质性1。尽管DLBCL患者的生存情况随着利妥昔单抗、蒽环类药物、干细胞移植以及嵌合抗原受体T细胞免疫疗法（CAR-T）等治疗手段的应用得到了极大改善，但是由于个体差异及耐药性，仍存在约40%复发难治患者2。此外，与DLBCL
