目的:C-C基序趋化因子受体7(CCR7)显著影响肿瘤的发生和进展,然而,它对肿瘤微环境(TME)和特定机制的影响仍然难以捉摸。炎性癌症相关成纤维细胞(iCAF),癌症相关成纤维细胞(CAF)的重要亚型,在调节TME和肿瘤生长中起关键作用,尽管潜在的分子机制还没有完全理解。本研究旨在确定CCR7是否参与iCAF的肿瘤调节,并阐明所涉及的具体机制。
方法:使用单细胞数据对CCR7敲除组和野生型组的CAF亚型进行差异基因分析。动物模型通过流式细胞术分选促进原代iCAF细胞的提取。通过qPCR和Western印迹检查DUSP1表达的变化以及慢病毒介导的敲低和过表达的效率。MOC1和MOC2细胞与iCAF共培养,随着随后对肿瘤细胞增殖变化的验证,迁移,使用CCK8,EdU,和伤口愈合试验。采用ELISA检测iCAF上清液中TGF-β1浓度的变化。
结果:CAF分为三种亚型-myCAF,iCAF,和基于单细胞数据的apCAF。分析显示,来自CCR7敲除组的iCAF中DUSP1表达显著增加,通过体外实验证实。将MOC1和MOC2细胞与显示慢病毒介导的DUSP1敲低的iCAF共培养导致抑制肿瘤细胞增殖,入侵,和移民。相比之下,与过表达DUSP1的iCAF共培养增强了这些能力。此外,在DUSP1敲低iCAF组中,上清液中的TGF-β1浓度增加,而在DUSP1过表达组下降。
结论:CCR7/DUSP1信号轴通过调节iCAF中的TGF-β1分泌来调节肿瘤生长。
OBJECTIVE: C-C motif chemokine receptor 7 (CCR7) significantly influences tumors onset and progression, yet its impact on the tumor microenvironment (TME) and specific mechanisms remain elusive. Inflammatory Cancer-Associated Fibroblasts (iCAF), a vital subtype of Cancer-Associated Fibroblasts (CAF), play a critical role in regulating the TME and tumor growth, though the underlying molecular mechanisms are not fully understood. This study aims to determine whether CCR7 participates in tumor regulation by iCAF and to elucidate the specific mechanisms involved.
METHODS: Differential gene analysis of CAF subtypes in CCR7 knockout and wild-type groups was conducted using single-cell data. Animal models facilitated the extraction of primary iCAF cells via flow cytometry sorting. Changes in
DUSP1 expression and the efficiency of lentivirus-mediated knockdown and overexpression were examined through qPCR and Western Blot. MOC1 and MOC2 cells were co-cultured with iCAF, with subsequent validation of changes in tumor cell proliferation, migration, and invasion using CCK8, EdU, and wound healing assays. ELISA was employed to detect changes in TGF-β1 concentration in the iCAF supernatant.
RESULTS: CAF was categorized into three subtypes-myCAF, iCAF, and apCAF-based on single-cell data. Analysis revealed a significant increase in DUSP1 expression in iCAF from the CCR7 knockout group, confirmed by in vitro experiments. Co-culturing MOC1 and MOC2 cells with iCAF exhibiting lentivirus-mediated
DUSP1 knockdown resulted in inhibited tumor cell proliferation, invasion, and migration. In contrast, co-culture with iCAF overexpressing
DUSP1 enhanced these capabilities. Additionally, the TGF-β1 concentration in the supernatant increased in the
DUSP1 knockdown iCAF group, whereas it decreased in the
DUSP1 overexpression group.
CONCLUSIONS: The CCR7/DUSP1 signaling axis regulates tumor growth by modulating TGF-β1 secretion in iCAF.