Gene expression profiling of peripheral blood B cells before and after surgery for hepatocellular carcinoma based on single-cell RNA sequencing technology
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 single-cell RNA sequencing(scRNA-seq)was performed.B cell number analysis,up-and down-regulated gene difference analysis,KEGG analysis,GSEA-KEGG and GSEA-GO 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 scRNA-seq to identify clusters of cells with marker genes MS4A1 and CD79A,namely,B-cell clusters.After regrouping analysis,the B-cell clusters were classified into three cell subpopulations,namely,memory B cells,naive B cells,and plasmablasts.Compared with pre-operative 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 up-regulated after surgery(P＜0.05),and 133 genes including RPS27A,RPS3A,RPS12 and RPS3 were significantly down-regulated after surgery(P＜0.05).KEGG analysis revealed that significantly down-regulated DEGs in post-operative B cells were mainly enriched in ribosomal,antigen processing and presentation pathways.GSEA-KEGG analysis revealed that the gene set up-regulated in post-operative B cells was significantly enriched in transcriptional dysregulation and P53 signaling pathway in cancer,while the gene set down-regulated was significantly enriched in ribosomal,antigen processing and presentation pathways.GSEA-GO analysis revealed that the gene set up-regulated in post-operative B cells was significantly enriched in cyclooxygenase P450 pathway,signaling pattern recognition receptor activity,and the down-regulated gene set was significantly enriched in immunoglobulin complex and T-cell receptor complex.Communication between B cells and T cells was enhanced after surgery,and the communication probability of post-operative ligand and receptor pairs of CD22-PTPRC between B cells and B cells,and between B cells and Mono cells was up-regulated(P＜0.01).Either before or after surgery,the CD22 signaling pathway had a strongly ability to regulate the communication between B cells and T cells.Conclusions The gene expression profile of peripheral blood B cells in HCC patients changes dynamically before and after surgery,and is correlated with the immune function of peripheral blood B cells.This result provides a new reference for the study of the anti-tumor immune function of peripheral blood B cells.
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