南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (9): 1267-1278.doi: 10.12122/j.issn.1673-4254.2022.09.01

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CD40LG是与浸润性乳腺癌肿瘤微环境中免疫和基质相关的预后标志物

郭 丽，马银玲，李 婷，李金平   

  1. 宁夏医科大学，宁夏 银川 750000；宁夏医科大学总医院，宁夏 银川 750000
  • 出版日期:2022-09-20 发布日期:2022-09-28

CD40LG is a novel immune- and stroma-related prognostic biomarker in the tumor microenvironment of invasive breast cancer

GUO Li, MA Yinling, LI Ting, LI Jinping   

  1. Ningxia Medical University, Yinchuan 750000, China; General Hospital of Ningxia Medical University, Yinchuan 750000, China
  • Online:2022-09-20 Published:2022-09-28

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摘要： 目的 旨在寻找与浸润性乳腺癌（BRCA）发生相关的肿瘤微环境（TME）相关基因，以预测其预后并为临床提供治疗靶点。方法 从癌症基因组图谱（TCGA）数据库中检索到RNA转录组数据和临床相关数据。利用ESTIMATE 算法计算基质评分和免疫评分。然后通过取交集筛选出差异表达基因（DEGs）。利用蛋白质-蛋白质相互作用（PPI）网络和单变量COX回归分析来确定DEGs中的核心基因。选取一个核心基因进行GSEA集富集分析和CIBERSORT分析，以分别区分核心基因表达的功能和肿瘤浸润免疫细胞（TICs）的比例。最终用Western blot和qRT-PCR对CD40LG的表达水平进行临床验证。结果 从TCGA中提取了1222个样本（124个正常样本和1098个肿瘤样本）进行分析，共获得了487个DEG。这些基因主要富集在与免疫相关的途径中。进一步的交叉分析揭示了11个关键基因，包括 CD40LG、ITK、CD5、CD3E、SPN、IL7R、CD48、CCL19、CD2、CD52和CD2711，这些基因被证明与乳腺癌TME状态相关。挑选了CD40LG进行进一步研究，结果表明，CD40LG高表达BRCA患者的总生存期（OS）比低表达BRCA患者更长（P=0.002），并且CD40LG表达在TNM 分期、肿瘤大小方面的差异也具有统计学意义（P<0.05）。GSEA 和 CIBERSORT 分析表明CD40LG的表达与TME中的免疫活性有关。Western blot和qRT-PCR结果显示CD40LG在乳腺癌细胞及癌组织中的蛋白及mRNA表达量均低于乳腺正常细胞及癌旁组织（P<0.05）。结论 CD40LG在TME中高表达与BRCA患者的生存呈正相关，所以CD40LG可能是一种新的用于预后预测的生物标志物。其生物学行为可能促进我们对肿瘤进展的分子机制和靶向治疗的理解。

关键词: 乳腺癌；肿瘤微环境；癌症基因组图谱；肿瘤浸润免疫细胞；CD40LG

Abstract: Objective To identify tumor microenvironment (TME)- related genes associated with the occurrence of invasive breast cancer as potential prognostic biomarkers and therapeutic targets. Methods RNA transcriptome data and clinically relevant data were retrieved from TCGA database, and the StromalScore and ImmuneScore were calculated using the ESTIMATE algorithm. The differentially expressed genes (DEGs) were screened by taking the intersection. A protein-protein interaction network was established, and univariate COX regression analysis was used to identify the core genes among the DEGs. A core gene was selected for GSEA and CIBERSORT analysis to determine the function of the core gene and the proportion of tumor-infiltrating immune cells , respectively. Western blotting and qRT-PCR were performed to verify the expression level of CD40LG in breast cancer cell lines and clinical specimens. Results A total of 1222 samples (124 normal and 1098 tumor samples) were extracted from TCGA for analysis, from which 487 DEGs were identified. These genes were mainly enriched in immune-related pathways, and crossover analysis identified 11 key genes (CD40LG, ITK, CD5, CD3E, SPN, IL7R, CD48, CCL19, CD2, CD52, and CD2711) associated with breast cancer TME status. CD40LG was selected as the core gene, whose high expression was found to be associated with a longer overall survival of breast cancer patients (P=0.002), and its expression level differed significantly with TNM stage and tumor size (P<0.05). GSEA and CIBERSORT analyses indicated that CD40LG expression level was associated with immune activity in the TME. Western blotting and qRT-PCR showed that the protein and mRNA expression of CD40LG were significantly lower in breast cancer cells and cancer tissues than in normal breast cells and adjacent tissues. Conclusions The high expression of CD40LG in TME is positively correlated with the survival of patients with invasive breast cancer, suggesting its value as a potential new biomarker for predicting prognosis of the patients.

Key words: breast cancer; tumor microenvironment; TCGA; tumor-infiltrating immune cells; CD40LG