单细胞 RNA 测序技术分析 CCR7 对小鼠口腔鳞状细胞癌微环境的影响。
Analysis of the effect of CCR7 on the microenvironment of mouse oral squamous cell carcinoma by single-cell RNA sequencing technology.
机构信息
Department of Oral Maxillofacial-Head and Neck Surgery, School and Hospital of Stomatology, China Medical University, Liaoning Provincial Key Laboratory of Oral Diseases, 117 Nanjing North Road, Heping District, Shenyang, Liaoning, 110002, People's Republic of China.
Department of Oral Biology, School of Dental Medicine, University at Buffalo, NY, Buffalo, 14214-8006, USA.
出版信息
J Exp Clin Cancer Res. 2024 Mar 27;43(1):94. doi: 10.1186/s13046-024-03013-y.
BACKGROUND
Studies have shown that CCR7, an important inflammatory factor, can promote the proliferation and metastasis of oral squamous cell carcinoma (OSCC), but its role in the tumor microenvironment (TME) remains unclear. This paper explores the role of CCR7 in the TME of OSCC.
METHODS
In this work, we constructed CCR7 gene knockout mice and OSCC mouse models. Single-cell RNA sequencing (scRNA-seq) and bioinformatics were used to analyze the differences in the OSCC microenvironment between three CCR7 gene knockout mice (KO) and three wild-type mice (WT). Immunohistochemistry, immunofluorescence staining, and flow cytometry were used to analyze the expression of key genes in significantly different cell types between the KO and WT groups. An in vitro experiment was used to verify the effect of CCR7 on M2 macrophage polarization.
RESULTS
In the mouse OSCC models, the tumor growth rate in the KO group was significantly lower than that in the WT group. Eight main cell types (including tumor cells, fibroblasts, macrophages, granulocytes, T cells, endothelial cells, monocytes, and B cells) were identified by Seurat analysis. The scRNA-seq results showed that the proportion of tumor cells was lower, but the proportion of inflammatory cells was significantly higher in the KO group than in the WT group. CellPhoneDB analysis results indicated a strong interaction relationship between tumor cells and macrophages, T cells, fibroblasts, and endothelial cells. Functional enrichment results indicated that the expression level of the Dusp1 gene in the KO group was generally higher than that in the WT group in various cell types. Macrophage subclustering results indicated that the proportion of M2 macrophages in the KO group was lower than that in the WT group. In vitro experimental results showed that CCR7 can promote M2 macrophage polarization, thus promoting the proliferation, invasion and migration of OSCC cells.
CONCLUSIONS
CCR7 gene knockout can significantly inhibit the growth of mouse oral squamous cell carcinoma by promoting the polarization of M2 macrophages.
背景
研究表明，CCR7 是一种重要的炎症因子，可促进口腔鳞状细胞癌（OSCC）的增殖和转移，但它在肿瘤微环境（TME）中的作用尚不清楚。本文探讨了 CCR7 在 OSCC 肿瘤微环境中的作用。
方法
在这项工作中，我们构建了 CCR7 基因敲除小鼠和 OSCC 小鼠模型。利用单细胞 RNA 测序（scRNA-seq）和生物信息学方法分析了 3 只 CCR7 基因敲除（KO）小鼠和 3 只野生型（WT）小鼠 OSCC 微环境的差异。免疫组织化学、免疫荧光染色和流式细胞术用于分析 KO 组和 WT 组中差异显著的细胞类型中的关键基因表达。体外实验用于验证 CCR7 对 M2 巨噬细胞极化的影响。
结果
在小鼠 OSCC 模型中，KO 组的肿瘤生长速度明显低于 WT 组。通过 Seurat 分析鉴定了 8 种主要细胞类型（包括肿瘤细胞、成纤维细胞、巨噬细胞、粒细胞、T 细胞、内皮细胞、单核细胞和 B 细胞）。scRNA-seq 结果显示，KO 组肿瘤细胞比例较低，但炎症细胞比例明显高于 WT 组。CellPhoneDB 分析结果表明肿瘤细胞与巨噬细胞、T 细胞、成纤维细胞和内皮细胞之间存在强烈的相互作用关系。功能富集结果表明，在各种细胞类型中，KO 组的 Dusp1 基因表达水平普遍高于 WT 组。巨噬细胞亚群聚类结果表明，KO 组 M2 型巨噬细胞的比例低于 WT 组。体外实验结果表明，CCR7 可促进 M2 型巨噬细胞极化，从而促进 OSCC 细胞的增殖、侵袭和迁移。
结论
CCR7 基因敲除可通过促进 M2 型巨噬细胞极化显著抑制小鼠口腔鳞状细胞癌的生长。
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