尽管联合免疫治疗和靶向治疗对于肝细胞癌（HCC）显示了积极的结果，但预后仍然较差。趋化因子及其受体在HCC的发展中起到重要作用，但其在HCC中的意义尚未完全阐明。本研究旨在建立与趋化因子相关的预后标志，并研究这些基因与肿瘤免疫微环境（TIME）的关联。从TCGA队列筛选了342例HCC患者。采用最小绝对收缩和选择算子回归和Cox比例风险回归分析建立了预后标志。使用ICGC数据库中的LIHC-JP队列进行了外部验证。来自GEO数据库的单细胞RNA测序（scRNA-seq）数据。建立了两个计分表来评估HCC患者的预后。采用RT-qPCR验证了预测标志中基因表达的差异。成功建立了包含两个趋化因子（CCL14，CCL20）和一个趋化因子受体（CCR3）的预后标志。根据中位数风险分数，将HCC患者分为高风险组和低风险组。发现低风险组的患者比高风险组的患者预后更好。单变量和多变量Cox回归分析的结果显示，该预后标志可被视为HCC患者预后的独立风险因素。我们发现高风险组与低风险组在各种免疫细胞亚型浸润、肿瘤突变负荷、生物通路、免疫激活或抑制基因的表达以及不同组对化疗药物和小分子靶向药物敏感性方面存在显著差异。随后的单细胞分析结果显示，CCL20的高表达与HCC转移相关。RT-qPCR结果表明，HCC与其相邻非肿瘤组织在CCL14、CCL20和CCR3的表达方面存在明显差异。本研究深入探讨了新型预后生物标记与TIME的关联，并进行了验证。这些结果在未来可应用于改善HCC的免疫治疗或靶向治疗的疗效。版权所有©2023张、毛、王、张、石、常、吴、张、许和鲁。

Despite encouraging results from immunotherapy combined with targeted therapy for hepatocellular carcinoma (HCC), the prognosis remains poor. Chemokines and their receptors are an essential component in the development of HCC, but their significance in HCC have not yet been fully elucidated. We aimed to establish chemokine-related prognostic signature and investigate the association between the genes and tumor immune microenvironment (TIME).342 HCC patients have screened from the TCGA cohort. A prognostic signature was developed using least absolute shrinkage and selection operator regression and Cox proportional risk regression analysis. External validation was performed using the LIHC-JP cohort deployed from the ICGC database. Single-cell RNA sequencing (scRNA-seq) data from the GEO database. Two nomograms were developed to estimate the outcome of HCC patients. RT-qPCR was used to validate the differences in the expression of genes contained in the signature.The prognostic signature containing two chemokines-(CCL14, CCL20) and one chemokine receptor-(CCR3) was successfully established. The HCC patients were stratified into high- and low-risk groups according to their median risk scores. We found that patients in the low-risk group had better outcomes than those in the high-risk group. The results of univariate and multivariate Cox regression analyses suggested that this prognostic signature could be considered an independent risk factor for the outcome of HCC patients. We discovered significant differences in the infiltration of various immune cell subtypes, tumor mutation burden, biological pathways, the expression of immune activation or suppression genes, and the sensitivity of different groups to chemotherapy agents and small molecule-targeted drugs in the high- and low-risk groups. Subsequently, single-cell analysis results showed that the higher expression of CCL20 was associated with HCC metastasis. The RT-qPCR results demonstrated remarkable discrepancies in the expression of CCL14, CCL20, and CCR3 between HCC and its paired adjacent non-tumor tissues.In this study, a novel prognostic biomarker explored in depth the association between the prognostic model and TIME was developed and verified. These results may be applied in the future to improve the efficacy of immunotherapy or targeted therapy for HCC.Copyright © 2023 Zhang, Mao, Wang, Zhang, Shi, Chang, Wu, Zhang, Xu and Lu.