Background/Objectives:Cancer-associated fibroblasts (CAFs) play a pivotal role in the tumor microenvironment. We conducted an analysis using RNA sequencing to identify specific markers for CAFs compared to normal fibroblasts (NFs) in non-small-cell carcinoma (NSCLC).Methods:CAFs and NFs were isolated and cultured from tumor tissues (primary tumor or metastatic lymph nodes) and matched non-tumor tissues, respectively. Bulk RNA sequencing was conducted on isolated CAFs and normal fibroblast NFs. Differential expressions, gene set enrichment, and CAF subpopulation prediction analyses were performed.Results:During the study period, 27 CAFs and 12 NFs were isolated and cultured from tumor and non-tumor tissues in patients with treatment-naïve NSCLC. Among them, 22 CAFs and 11 NFs were included in the RNA sequencing analysis. The 22 CAF samples consisted of 12 adenocarcinomas and 10 squamous cell carcinomas (SqCC), with 16 samples from the lungs and 6 samples from the lymph nodes. Notably,COL11A1,GREM1,CD36, andGAS6showed a higher expression in CAFs than in NFs, whereasTNCandCXCL2were more abundantly expressed in NFs.CD36levels were elevated in CAFs from lymph nodes (LN-CAFs) compared with those from lung specimens (Lung-CAFs) and NFs.COL11A1levels in Lung-CAFs surpassed those in LN-CAFs and NFs. BothGREM1andGAS6showed a strong expression in Lung-CAFs and LN-CAFs relative to NFs. CAFs exhibited features of the myofibroblast CAF subpopulation, whereas NFs displayed traits of the antigen-presenting CAF subtype. In the co-culture model of CAFs and THP-1 cells, the knockdown ofGREM1orGAS6in CAFs significantly decreased the M2 marker expression in macrophages.Conclusions:In NSCLC,GREM1andGAS6can be valuable diagnostic targets for CAFs from primary tumors and metastatic sites; they warrant further study.
背景/目的：癌相关成纤维细胞（CAFs）在肿瘤微环境中扮演关键角色。本研究通过RNA测序分析，旨在识别非小细胞肺癌（NSCLC）中CAFs相较于正常成纤维细胞（NFs）的特异性标志物。 方法：分别从肿瘤组织（原发肿瘤或转移淋巴结）及匹配的非肿瘤组织中分离培养CAFs和NFs。对分离出的CAFs和NFs进行批量RNA测序，并开展差异表达分析、基因集富集分析及CAF亚群预测分析。 结果：在研究期间，从初治NSCLC患者的肿瘤及非肿瘤组织中成功分离培养了27例CAFs和12例NFs。其中22例CAFs和11例NFs纳入RNA测序分析。22例CAFs样本包括12例腺癌和10例鳞状细胞癌，其中16例来源于肺组织，6例来源于淋巴结。值得注意的是，COL11A1、GREM1、CD36和GAS6在CAFs中的表达高于NFs，而TNC和CXCL2在NFs中表达更丰富。与肺组织来源CAFs（Lung-CAFs）和NFs相比，淋巴结来源CAFs（LN-CAFs）中CD36水平升高；Lung-CAFs中COL11A1水平则高于LN-CAFs和NFs。GREM1和GAS6在Lung-CAFs和LN-CAFs中均较NFs呈现强表达。CAFs表现出肌成纤维细胞CAF亚群特征，而NFs则显示抗原呈递CAF亚型特性。在CAFs与THP-1细胞的共培养模型中，敲低CAFs中的GREM1或GAS6可显著降低巨噬细胞的M2标志物表达。 结论：在NSCLC中，GREM1和GAS6可作为原发肿瘤及转移部位CAFs的重要诊断靶标，值得进一步深入研究。
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