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中华肝脏外科手术学电子杂志

中华肝脏外科手术学电子杂志 ›› 2025, Vol. 14 ›› Issue (04) : 609 -618. doi: 10.3877/cma.j.issn.2095-3232.2025.04.017

基础研究

多组学分析HAPLN1与肝癌预后及免疫细胞浸润关系
刘晓萍1,2, 汪嵘嵘1,2, 吴佳慧1, 吴紫云1,2, 周伯宣1,2,3,†()   
  1. 1330006　南昌大学第一附属医院乳腺专病中心
    2330006　南昌大学第一附属医院南昌大学第一临床医学院
    3330006　南昌大学第一附属医院输血医学江西省重点实验室
  • 收稿日期:2025-03-06 出版日期:2025-08-10
  • 通信作者: 周伯宣
  • 基金资助:
    江西省自然科学基金青年项目(20242BAB20404); 江西省教育厅重点项目(GJJ2400104); 南昌大学第一附属医院青年人才科研培养基金(YFYPY2023103); 南昌大学第一附属医院专职人才引进与科研启动基金(RSC-0020)

Multiomic analysis of the relationship between HAPLN1 and prognosis of hepatocellular carcinoma and immune cell infiltration

Xiaoping Liu1,2, Rongrong Wang1,2, Jiahui Wu1, Ziyun Wu1,2, Boxuan Zhou1,2,3,†()   

  1. 1Center for Breast Diseases, the First Affiliated Hospital of Nanchang University, Nanchang 330006, China
    2The First Clinical Medical College of Nanchang University, the First Affiliated Hospital of Nanchang University, Nanchang 330006, China
    3Jiangxi Provincial Key Laboratory of Blood Transfusion Medicine, the First Affiliated Hospital of Nanchang University, Nanchang 330006, China
  • Received:2025-03-06 Published:2025-08-10
  • Corresponding author: Boxuan Zhou
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引用本文：

刘晓萍, 汪嵘嵘, 吴佳慧, 吴紫云, 周伯宣. 多组学分析HAPLN1与肝癌预后及免疫细胞浸润关系[J/OL]. 中华肝脏外科手术学电子杂志, 2025, 14(04): 609-618.

Xiaoping Liu, Rongrong Wang, Jiahui Wu, Ziyun Wu, Boxuan Zhou. Multiomic analysis of the relationship between HAPLN1 and prognosis of hepatocellular carcinoma and immune cell infiltration[J/OL]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2025, 14(04): 609-618.

目的

分析透明质酸和蛋白聚糖连接蛋白1（HAPLN1）表达与肝细胞癌（肝癌）患者预后的关系，并探讨其与肝癌免疫细胞浸润的关系。

方法

基于TCGA数据库分析HAPLN1在肝癌中的表达及其与临床病理特征、预后的关系。通过Cox单因素和多因素分析筛选与肝癌预后相关的危险因素。构建列线图模型对生存预后进行量化分析，ROC曲线评估HAPLN1对肝癌患者预后的预测价值。采用Cluster Profile软件包对肝癌HAPLN1高、低表达组的差异表达基因对应的通路和细胞功能进行基因本体论分析（GO）和京都基因与基因组百科全书（KEGG）分析。利用基因集富集分析‌（GSEA）对HAPLN1相关通路进行可视化。采用CIBERSORT核心算法评估肝癌中HAPLN1与24种免疫细胞的相关性，采用ssGSEA算法评估24种免疫细胞在HAPLN1高、低表达组肝癌中的富集情况，进一步探讨肝癌中免疫检查点与HAPLN1的相关性。

结果

TCGA数据库分析显示，HAPLN1在肝癌组织中上调，且与配对正常肝组织相比，肝癌组织的HAPLN1表达明显高于正常肝组织（P=0.002），HAPLN1高表达的肝癌患者总生存期更低（HR=1.560，95%CI：1.100~2.218;P=0.013）。HAPLN1表达与T分期、血管侵犯和病理分级显著相关（χ2=9.149，9.518，6.614；P<0.05）。Cox多因素回归分析显示，T分期（HR=2.622，95%CI：1.692~4.062；P<0.001)和HAPLN1表达（HR=1.622，95%CI：1.072~2.585；P=0.023）是肝癌患者预后的独立危险因素。预后列线图分析显示，HAPLN1对于肝癌预后具有较好的预测价值（AUC=0.737，95%CI：0.685~0.788）。GO和KEGG富集分析显示，HAPLN1的差异表达基因主要富集于顶部质膜、信号受体激活和信号配体活性功能以及胆汁分泌等功能通路；免疫细胞相关性分析显示，HAPLN1与多种免疫细胞密切相关，且与多种免疫检查点呈正相关。

结论

HAPLN1高表达的肝癌患者预后更差，基于HAPLN1表达联合病理TNM分期构建的列线图模型为预测肝癌患者预后提供了有效参考。HAPLN1可能通过调控肿瘤免疫微环境促进肝癌进展。

关键词: 癌，肝细胞, 透明质酸和蛋白聚糖连接蛋白1, 预后, 模型, 富集分析, 免疫检查点
Objective

To analyze the relationship between the expression level of hyaluronan and proteoglycan link protein 1 (HAPLN1) and the prognosis of hepatocellular carcinoma (HCC) patients, and to investigate its correlation with immune cell infiltration of HCC.

Methods

Based on the TCGA database, the expression of HAPLN1 in HCC and its relationship with clinicopathological features and prognosis were analyzed. The risk factors of prognosis of HCC were screened by univariate and multivariate Cox’s regression analyses. The nomogram model was constructed to quantitatively analyze the survival and prognosis. ROC curve was delineated to evaluate the predictive value of HAPLN1 for clinical prognosis of HCC patients. The pathway and cell function of differentially-expressed genes between the high and low HAPLN1 expression groups were subject to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses using Cluster Profile software package for HAPLN1-related pathways were visualized by using Gene Set Enrichment Analysis (GSEA). The correlation between HAPLN1 and 24 types of immune cells in HCC was analyzed by CIBERSORT core algorithm. The enrichment of 24 types of immune cells in HCC in high and low HAPLN1 expression groups was assessed by ssGSEA algorithm. The correlation between immune checkpoints and HAPLN1 in HCC was further investigated.

Results

TCGA database analysis showed that the expression level of HAPLN1 was up-regulated in HCC tissues, significantly higher than that in matched normal liver tissues (P=0.002). HCC patients with high expression of HAPLN1 had shorter overall survival (HR=1.560, 95%CI: 1.100-2.218, P=0.013). HAPLN1 expression was significantly correlated with T stage, vascular invasion and pathological stage (χ2=9.149, 9.518, 6.614; all P<0.05). Multivariate Cox’s regression analysis showed that T stage (HR=2.622, 95%CI: 1.692-4.062; P<0.05) and HAPLN1 expression (HR=1.622, 95%CI: 1.072~2.585; P=0.023) were the independent risk factors for clinical prognosis of HCC patients. Prognostic nomogram analysis revealed that HAPLN1 had high predictive value for clinical prognosis of HCC (AUC=0.737, 95%CI: 0.685-0.788). GO and KEGG enrichment analyses showed that the differentially-expressed genes of HAPLN1 were mainly enriched in the functional pathways such as apical plasma membrane, signal receptor activation and signal ligand activity and bile secretion, etc. Immune cell correlation analysis showed that HAPLN1 was closely associated with multiple immune cells in HCC, and was positively correlated with immune checkpoints.

Conclusions

HCC patients with high expression of HAPLN1 have poor prognosis. The nomogram model based on the expression of HAPLN1 combined with pathological TNM staging provides effective reference for predicting clinical prognosis of HCC patients. HAPLN1 may promote the progression of HCC through regulating tumor immune microenvironment.

Key words: Carcinoma,hepatocellular, Hyaluronan and proteoglycan link protein 1 (HAPLN1), Prognosis, Model, Enrichment analysis, Immune checkpoint
图1 根据TCGA数据库分析HAPLN1在泛癌和肝癌中表达水平注：a为肝癌中2 426个差异基因表达的火山图，上调的基因标为红色，下调的基因标为蓝色；b为HAPLN1在肝癌和癌旁正常肝组织中表达差异的配对样本图；c为HAPLN1高、低表达组的总生存期预后分析；HAPLN1为透明质酸和蛋白聚糖连接蛋白1
表1 肝癌患者HAPLN1表达水平与临床病理特征的关系[例(%)]
临床病理特征 HAPLN1 χ 2值 P值
低表达组 高表达组
年龄     0.781 0.377
≤60岁 93 (24.9) 84 (22.5)    
>60岁 94 (25.2) 102 (27.3)    
性别     1.478 0.224
女 55 (14.7) 66 (17.6)    
男 132 (35.3) 121 (32.4)    
T分期     9.149 0.002
T1~T2 152 (41.0) 126 (34.0)    
T3~T4 34 (9.2) 59 (15.9)    
N1分期 3 (1.2) 1 (0.4) 0.237 0.625
M1分期 2 (0.7) 2 (0.7) 0.225 >0.999
AFP>400 μg/L 34 (12.1) 31 (11.1) 0.090 0.765
血管侵犯 44 (13.8) 66 (20.8) 9.518 0.002
病理分级     6.615 0.010
Ⅰ~Ⅱ 142 (40.6) 118 (33.7)    
Ⅲ~Ⅳ 35 (10.0) 55 (15.7)    
表2 肝癌患者临床病理参数的单因素和多因素分析
参数 单因素分析 多因素分析
HR(95%CI) P值 HR(95%CI) P值
年龄>60岁 1.205 (0.850~1.708) 0.295    
男性 0.793(0.557~1.130) 0.200    
T3~T4期 2.598 (1.826~3.697) <0.001 2.622 (1.692~4.062) <0.001
N1期 2.029 (0.497~8.281) 0.324    
M1期 4.077 (1.281~12.973) 0.017 2.281 (0.697~7.463) 0.173
无血管侵犯 1.344 (0.887~2.035) 0.163    
HAPLN1高表达 1.631 (1.147~2.319) 0.006 1.665 (1.072~2.585) 0.023
图2 构建肝癌预后列线图模型并验证HAPLN1表达在肝癌中的诊断价值注：a为肝癌患者1、3、5年总生存影响因素的列线图模型；b为列线图模型的Calibration校准曲线；c为ROC曲线；HAPLN1为透明质酸和蛋白聚糖连接蛋白1
图3 肝癌HAPLN1基因共表达分析注：a为与HAPLN1正负相关性最强的12个基因共表达和弦图；b为HAPLN1与两个最强正相关基因ANGPT2、HOXD9及两个最强负相关基因FTCD、SLC22A1的相关性散点图；HAPLN1为透明质酸和蛋白聚糖连接蛋白1
图4 肝癌中HAPLN1差异表达基因分析及其通路和细胞功能鉴定注：a为1 024个肝癌差异表达基因的GO（左）、KEGG（右）富集情况；b为GSEA通路富集（左）分析和功能富集分析（右）；HAPLN1为透明质酸和蛋白聚糖连接蛋白1，GO为基因本体，‌GSEA为基因集富集分析
图5 肝癌中HAPLN1表达与免疫细胞浸润及免疫检查点基因的相关性分析注：a为HAPLN1与24种免疫细胞的相关性棒棒图，*为P<0.05，**为P<0.01，***为P<0.001，红色代表正相关，蓝色代表负相关，颜色越深或棒状直径越大表示相关性越强；b为肝癌中HAPLN1高表达组与低表达组的24种免疫细胞富集情况对比；c为肝癌中与HAPLN1相关性最强的前15个免疫检查点基因的相关系数热图；HAPLN1为透明质酸和蛋白聚糖连接蛋白1
[1]
应倩, 汪媛. 肝癌流行现况和趋势分析[J]. 中国肿瘤, 2020, 29(3): 185-191. DOI: 10.11735/j.issn.1004-0242.2020.03.A005.
[2]
谢林虎, 王凤玲, 刘建军. 原发性肝癌介入联合免疫治疗的研究进展[J]. 长春中医药大学学报, 2024, 40(5): 586-590. DOI: 10.13463/j.cnki.cczyy.2024.05.026.
[3]
周红, 刘永刚, 张海博, 等. 肿瘤相关成纤维细胞在肝细胞癌中的作用及机制[J]. 胃肠病学和肝病学杂志, 2023, 32(11): 1304-1310. DOI: 10.3969/j.issn.1006-5709.2023.11.021.
[4]
符赞美, 蔡伟国, 龙清云, 等.基于生物信息学分析DNASE1L3在肝细胞癌中表达及对预后的影响[J]. 武汉大学学报(医学版), 2024, 45(8): 939-945.DOI:10.14188/j.1671-8852.2023.0458.
[5]
杨飞龙, 洪锴, 赵国江, 等. 基于长链非编码RNA的生物信息学分析构建膀胱癌预后模型并确定预后生物标志物[J]. 北京大学学报(医学版), 2019, 51(4): 615-622. DOI: 10.19723/j.issn.1671-167X.2019.04.003.
[6]
Wiedmann L, De Angelis Rigotti F, Vaquero-Siguero N, et al. HAPLN1 potentiates peritoneal metastasis in pancreatic cancer[J]. Nat Commun, 2023, 14(1): 2353. DOI: 10.1038/s41467-023-38064-w.
[7]
Zhang T, Li X, He Y, et al. Cancer-associated fibroblasts-derived HAPLN1 promotes tumour invasion through extracellular matrix remodeling in gastric cancer[J]. Gastric Cancer, 2022, 25(2): 346-359. DOI: 10.1007/s10120-021-01259-5.
[8]
丁亿, 黄浩轩, 熊世达, 等. 透明质酸和蛋白聚糖连接蛋白家族在肿瘤中的研究进展[J]. 南昌大学学报(医学版), 2022, 62(6): 92-96. DOI: 10.13764/j.cnki.ncdm.2022.06.018.
[9]
De Bakshi D, Chen YC, Wuerzberger-Davis SM, et al. Ectopic CH60 mediates HAPLN1-induced cell survival signaling in multiple myeloma[J]. Life Sci Alliance, 2022, 6(3): e202201636. DOI: 10.26508/lsa.202201636.
[10]
Ecker BL, Kaur A, Douglass SM, et al. Age-related changes in HAPLN1 increase lymphatic permeability and affect routes of melanoma metastasis[J]. Cancer Discov,2019,9(1):82-95. DOI: 10.1158/2159-8290.CD-18-0168.
[11]
Liu J, Lichtenberg T, Hoadley KA, et al. An integrated TCGA pan-cancer clinical data resource to drive high-quality survival outcome analytics[J]. Cell, 2018, 173(2): 400-416.e11. DOI: 10.1016/j.cell.2018.02.052.
[12]
吴爽, 张涛元, 李俏, 等. MED8表达与肝癌临床病理特征及其预后的相关性分析[J]. 实用癌症杂志, 2024, 39(3): 411-416.
[13]
常晴, 刘佳, 曲爱林, 等. 利用数据库信息分析NAMPT与肝癌的病理特征和免疫浸润的关联性[J]. 山东大学学报(医学版), 2023, 61(4): 26-36. DOI: 10.6040/j.issn.1671-7554.0.2022.0913.
[14]
周静, 王心悦, 李兆娜, 等. 肺腺癌自噬相关基因预后风险评分模型构建及验证[J]. 中国肺癌杂志, 2021, 24(8): 557-566. DOI: 10.3779/j.issn.1009-3419.2021.103.09.
[15]
王文博, 黄建钊, 刘延. 原发性肝癌的手术与非手术治疗方法研究进展[J]. 中国实用医药, 2024, 19(12): 175-177. DOI: 10.14163/j.cnki.11-5547/r.2024.12.048.
[16]
Zhou D, Jang JM, Yang G, et al. A novel role of hyaluronic acid and proteoglycan link protein 1 (HAPLN1) in delaying vascular endothelial cell senescence[J]. Biomol Ther (Seoul), 2023, 31(6): 629-639. DOI: 10.4062/biomolther.2023.096.
[17]
LeBleu VS, Neilson EG. Origin and functional heterogeneity of fibroblasts[J]. FASEB J, 2020, 34(3): 3519-3536. DOI: 10.1096/fj.201903188r.
[18]
Wang Y, Xu X, Marshall JE, et al. Loss of hyaluronan and proteoglycan link protein-1 induces tumorigenesis in colorectal cancer[J]. Front Oncol, 2021, 11: 754240. DOI: 10.3389/fonc.2021.754240.
[19]
贺红柳, 崔美英, 王丰, 等. HAPLN1通过IKK/p65信号通路诱导大肠癌HT-29细胞产生MTX耐药[J]. 中国病理生理杂志, 2019, 35(4): 621-627. DOI: 10.3969/j.issn.1000-4718.2019.04.007.
[20]
韩朝辉, 龙静, 刘洁玲, 等. miR-199 b-5p调控HAPLN1对宫颈癌细胞顺铂耐药性的影响[J]. 河北医药, 2022, 44(19): 2896-2901. DOI: 10.3969/j.issn.1002-7386.2022.19.003.
[21]
Mebarki S, Désert R, Sulpice L, et al. De novo HAPLN1 expression hallmarks Wnt-induced stem cell and fibrogenic networks leading to aggressive human hepatocellular carcinomas[J]. Oncotarget, 2016, 7(26): 39026-39043. DOI: 10.18632/oncotarget.9346.
[22]
Derynck R, Muthusamy BP, Saeteurn KY. Signaling pathway cooperation in TGF-β-induced epithelial-mesenchymal transition[J]. Curr Opin Cell Biol,2014,31:56-66. DOI: 10.1016/j.ceb.2014.09.001.
[23]
Liu Y, Zhou R, Yuan X, et al. DACH1 is a novel predictive and prognostic biomarker in hepatocellular carcinoma as a negative regulator of Wnt/β-catenin signaling[J]. Oncotarget, 2015, 6(11): 8621-8634. DOI: 10.18632/oncotarget.3281.
[24]
Kitisin K, Ganesan N, Tang Y, et al. Disruption of transforming growth factor-beta signaling through beta-spectrin ELF leads to hepatocellular cancer through cyclin D1 activation[J]. Oncogene, 2007, 26(50): 7103-7110. DOI: 10.1038/sj.onc.1210513.
[25]
Zhang K, Zhang M, Luo Z, et al. The dichotomous role of TGF-β in controlling liver cancer cell survival and proliferation[J]. J Genet Genomics, 2020, 47(9): 497-512. DOI: 10.1016/j.jgg.2020.09.005.
[26]
张岩岩, 曹静, 陈晓彤, 等. 肝组织特异性基因IGFALS、CYP3A4、SLC22A1和CYP2E1可能与肝癌预后不良相关[J]. 中国医学科学院学报, 2021, 43(3): 371-381. DOI: 10.3881/j.issn.1000-503X.13096.
[27]
Liu Z, Li Y, Liu Y, et al. Expression and clinical significance of BDH1 in liver cancer[J]. Medicine, 2021, 100(48): e28013. DOI: 10.1097/MD.0000000000028013.
[28]
刘晓丽, 谭钦文, 徐健, 等. 氨基酸代谢重编程对肝细胞癌免疫微环境的影响[J].临床肝胆病杂志,2024, 40(12): 2531-2537. DOI: 10.12449/JCH241226.
[29]
沈波, 聂玉强. 过氧化物酶体增殖物激活受体γ抑制肝癌细胞的增殖生长和侵袭转移[J]. 临床肝胆病杂志, 2013, 29(9): 702-706. DOI: 10.3969/j.issn.1001-5256.2013.09.018.
[30]
祁婉婉, 何奕婷, 张慧林. 子宫内膜癌免疫微环境的研究进展[J]. 现代妇产科进展, 2025, 34(1): 70-74. DOI: 10.13283/j.cnki.xdfckjz.2025.01.032.
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