基于单细胞RNA测序技术的肝癌手术前后外周血B细胞基因表达谱研究  

Gene expression profiling of peripheral blood B cells before and after surgery for hepatocellular carcinoma based on single-cell RNA sequencing technology

作　　者：梁思 盛勇红 刘顺[2] 刘美良 杨钰 雷蕾 巫丽丽[4] 曾小云[1] LIANG Si;SHENG Yonghong;LIU Shun;LIU Meiliang;YANG Yu;LEI Lei;WU Lili;ZENG Xiaoyun(Department of Epidemiology and Health Statistics,School of Public Health,Guangxi Medical University;Department of Child and Adolescent Health&Maternal and Child Health,School of Public Health,Guangxi Medical University;Department of Clinical Internal Medicine Nursing,School of Nursing,Guangxi Medical University;Department of Endocrinology and Metabolic Nephrology,Guangxi Medical University Cancer Hospital,Nanning 530021,China)

机构地区：[1]广西医科大学公共卫生学院流行病与卫生统计学教研室,南宁530021 [2]广西医科大学公共卫生学院儿少卫生与妇幼保健医学教研室,南宁530021 [3]广西医科大学护理学院临床内科护理学教研室,南宁530021 [4]广西医科大学附属肿瘤医院内分泌代谢肾病内科,南宁530021

出　　处：《中国癌症防治杂志》2024年第4期433-442,共10页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：国家自然科学基金项目(82060616)。

摘　　要：目的探索肝细胞癌(hepatocellular carcinoma,HCC)患者手术前后外周血B细胞基因表达谱的变化。方法收集2022年9月在广西医科大学附属肿瘤医院行肝切除术治疗的3例HCC患者手术前及手术后的外周血样本,提取外周血单个核细胞(peripheral blood mononuclear cell,PBMC)并进行单细胞RNA测序(single-cell RNA sequencing,scRNA-seq)。根据测序数据进行B细胞数量分析、上下调基因差异分析、KEGG分析、GSEA-KEGG及GSEA-GO分析和细胞通讯分析,比较手术前后B细胞的动态改变。结果PBMC经scRNA-seq分析识别出了标记基因为MS4A1、CD79A的细胞簇,即B细胞群。B细胞群重新聚类分析后可分为记忆B(memory B)细胞、初始B(naive B)细胞和浆母细胞(plasmablasts)共3个细胞亚群。与术前组相比,术后组naive B和plasmablasts细胞占比升高,memory B细胞占比降低。手术前后B细胞中共筛选出285个显著差异表达基因(differentially expressed genes,DEGs),其中S100A8、S100A9、IL1R2、MALAT1等152个基因在手术后显著上调(P<0.05),RPS27A、RPS3A、RPS12、RPS3等133个基因在手术后显著下调(P<0.05)。KEGG分析发现手术后B细胞中显著下调的DEGs主要富集到核糖体、抗原处理和提呈通路。GSEA-KEGG分析发现手术后B细胞中表达上调的基因集显著富集于癌症中的转录失调、P53信号通路,表达下调的基因集显著富集于核糖体、抗原处理和提呈通路;GSEA-GO分析发现手术后B细胞中表达上调的基因集显著富集于环氧化酶P450途径、信号模式识别受体活性,表达下调的基因集显著富集于免疫球蛋白复合物、T细胞受体复合物。手术后B细胞与T细胞之间的通讯增强,且手术后配体-受体对CD22-PTPRC在B细胞与B细胞、B细胞与Mono细胞之间的通讯概率上调(P<0.01)。无论是手术前还是手术后,CD22信号通路对B细胞与T细胞之间的通讯均具有较强调控能力。结论HCC患者外周血B细胞基因表达谱在手术前Objective To investigate the changes of gene expression profiles of peripheral blood B cells in hepatocellular carcinoma(HCC)patients before and after surgery.Methods Peripheral blood samples of 3 HCC patients who underwent hepatectomy for treatment at the Guangxi Medical University Cancer Hospital in September 2022 were collected before and after surgery,peripheral blood monoculear cell(PBMC)were extracted and singlecell RNA sequencing(scRNAseq)was performed.B cell number analysis,upand downregulated gene difference analysis,KEGG analysis,GSEAKEGG and GSEAGO analysis,and cell communication analysis were performed based on the sequencing data to compare the dynamic changes of B cells before and after surgery.Results PBMC was analyzed by scRNAseq to identify clusters of cells with marker genes MS4A1 and CD79A,namely,Bcell clusters.After regrouping analysis,the Bcell clusters were classified into three cell subpopulations,namely,memory B cells,naive B cells,and plasmablasts.Compared with preoperative group,the percentage of naive B cells and plasmablasts increased,and the percentage of memory B cells decreased after operation.A total of 285 significantly differentially expressed genes(DEGs)were screened in B cells before and after surgery,of which 152 genes including S100A8,S100A9,IL1R2 and MALAT1 were significantly upregulated after surgery(P<0.05),and 133 genes including RPS27A,RPS3A,RPS12 and RPS3 were significantly downregulated after surgery(P<0.05).KEGG analysis revealed that significantly downregulated DEGs in postoperative B cells were mainly enriched in ribosomal,antigen processing and presentation pathways.GSEAKEGG analysis revealed that the gene set upregulated in postoperative B cells was significantly enriched in transcriptional dysregulation and P53 signaling pathway in cancer,while the gene set downregulated was significantly enriched in ribosomal,antigen processing and presentation pathways.GSEAGO analysis revealed that the gene set upregulated in postoperative B cells was significantly enriched in

关 键 词：单细胞RNA测序 B细胞 肝癌 基因表达谱 

分 类 号：R735.7[医药卫生—肿瘤]