赖氨酰羟化酶2对肝细胞癌患者预后及肿瘤免疫浸润的影响  

Prognostic value of lysyl hydroxylase 2 in hepatocellular carcinoma and its impact on tumor immune infiltration

作　　者：王伟 周瑶 应万涛 孙薇 邢美宁 WANG Wei;ZHOU Yao;YING Wantao;SUN Wei;XING Meining(State Key Laboratory of Medical Proteomics,Beijing Proteome Research Center,National Center for Protein Sciences(Beijing),Institute of Lifeomics,Academy of Military Medical Sciences,Beijing 102206,China)

机构地区：[1]军事科学院军事医学研究院生命组学研究所,国家蛋白质科学中心(北京),北京蛋白质组研究中心,医学蛋白质组全国重点实验室,北京102206

出　　处：《癌变.畸变.突变》2025年第2期96-104,共9页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金(81903017);重点实验室自主研究课题(SKLP-K201901)。

摘　　要：目的:探究赖氨酰羟化酶2(PLOD2)在肝细胞癌(HCC)中的生物学功能,及其对患者预后和肿瘤免疫浸润的影响。方法:通过癌症基因组图谱(TCGA)数据库分析PLOD2在HCC中的表达,统计分析评估PLOD2表达与HCC临床病理特征之间的相关性及其预后价值,基因集富集分析(GSEA)揭示与PLOD2表达相关的信号通路。通过慢病毒介导的shRNA技术构建稳定敲低PLOD2的人肝癌细胞Huh7细胞系,并用Western blot法验证敲低效果。用CCK-8法、Transwell小室法和划痕实验验证PLOD2对Huh7细胞生物学行为的影响。用单样本基因集富集分析(ssGSEA)研究PLOD2表达与HCC免疫浸润之间的相关性。结果:生物信息学分析结果显示,与正常肝组织比较,PLOD2在HCC中显著高表达(P<0.05或P<0.01),并与患者年龄、T分期、病理分期、肿瘤状态、组织学分级和AFP水平呈正相关(P<0.05)。生存分析显示,PLOD2高表达组的中位生存时间均短于低表达组(P<0.05),PLOD2诊断HCC的受试者工作特征曲线下面积为0.656,提示PLOD2在HCC的诊断和预后预测中存在参考价值。此外,GSEA揭示了PLOD2高表达相关的信号通路包括癌通路、细胞周期、黏着斑等。细胞转染实验表明,与正常对照组细胞相比,敲低PLOD2能显著抑制Huh7细胞的增殖、侵袭和迁移能力(P<0.05),在敲低PLOD2的Huh7细胞中恢复表达PLOD2后细胞的迁移能力显著回升(P<0.05)。PLOD2高表达与HCC中Th2细胞浸润水平呈正相关(r=0.372,P<0.01),而与Th17和pDC等细胞免疫浸润水平呈负相关(r=-0.310,P<0.01)。结论:本研究揭示了PLOD2在HCC进展中的重要作用,PLOD2高表达与HCC患者的不良预后相关,可能通过调节免疫细胞的浸润和功能,在免疫抑制微环境中发挥关键作用。OBJECTIVE:To investigate biological function and prognostic value of lysyl hydroxylase 2(PLOD2)in hepatocellular carcinoma(HCC),as well as its impact on tumor immune infiltration.METHODS:PLOD2 expression in HCC was analyzed using data from The Cancer Genome Atlas(TCGA)database.Correlations between PLOD2 expression and clinicopathological characteristics and its prognostic value were evaluated by statistical analysis.Signaling pathways associated with PLOD2 expression were identified using gene set enrichment analysis(GSEA).A stable knockdown of PLOD2 in a human hepatocellular carcinoma cell line,Huh7,was achieved using lentivirus-mediated shRNA technology,and the knockdown effect was verified by Western blot.Effects of PLOD2 on biological behavior of Huh7 cells were assessed using CCK-8,Transwell,and scratch assays.Correlations between PLOD2 expression and HCC immune infiltration was investigated using single-sample gene set enrichment analysis(ssGSEA).RESULTS:Bioinformatics analysis revealed that PLOD2 was significantly overexpressed in HCC compared to normal liver tissues(P<0.05 or P<0.01)and was positively correlated with patient age,T stage,pathological stage,tumor status,histological grade,and AFP level(P<0.05).Survival analysis showed that the median survival time of the PLOD2 high-expression group was significantly shorter than that of the low-expression group(P<0.05).The area under the receiver operating characteristic(ROC)curve for PLOD2 in diagnosing HCC was 0.656,indicating that PLOD2 had reference value in the diagnosis and prognosis prediction of HCC.GSEA revealed that several signaling pathways were associated with high PLOD2 expression:cancer pathways,cell cycle,and adhesion junctions.Cell line validation experiments demonstrated that knockdown of PLOD2 significantly inhibited proliferation,invasion,and migration of Huh7 cells compared to control cells(P<0.05).Migration ability of Huh7 cells with PLOD2 knockdown was significantly reversed upon re-expression of PLOD2(P<0.05).Expression of PLOD2 was

关 键 词：赖氨酰羟化酶2 肝细胞癌 临床价值 免疫浸润 

分 类 号：R735.7[医药卫生—肿瘤]