Cancer-associated fibroblasts in gastric cancer affect malignant progression via the CXCL12-CXCR4 axis

Yan Qin; Fang Wang; Hengli Ni; Yao Liu; Yuan Yin; Xinyi Zhou; Guihua Gao; Qing Li; Xiaowei Qi; Jianming Li

doi:10.7150/jca.49707

Cancer-associated fibroblasts in gastric cancer affect malignant progression via the CXCL12-CXCR4 axis

J Cancer. 2021 Mar 19;12(10):3011-3023. doi: 10.7150/jca.49707. eCollection 2021.

Authors

Yan Qin^{1

2}, Fang Wang³, Hengli Ni¹, Yao Liu¹, Yuan Yin⁴, Xinyi Zhou^{2

3}, Guihua Gao², Qing Li⁵, Xiaowei Qi^{2

3}, Jianming Li¹

Affiliations

¹ Department of Pathology, Medical College of Soochow University, Soochow University, Suzhou, China.
² Department of Pathology, the Affiliated Hospital of Jiangnan University, Wuxi, China.
³ Jiangnan University School of Medicine, Wuxi, China.
⁴ Wuxi Oncology Institute, the Affiliated Hospital of Jiangnan University, Wuxi China.
⁵ Department of Oncology, The Second Affiliated Hospital of Soochow University Suzhou, China.

Abstract

Background: Cancer-associated fibroblasts (CAFs) are principal constituents of the tumor microenvironment (TME) and play a critical role in tumor progression. The CXCL12/CXCR4 axis regulates multiple facets of the TME. The aim of this study was to determine the relationship between CXCL12 expression in CAFs and the malignant progression of gastric cancer (GC). Methods: In the GEO (Gene Expression Omnibus) database, we performed transcriptome analysis on paired gastric cancer RNA sequencing samples, and scRNA analysis was performed on advanced malignant GC samples from the scRNA sequencing data set. Fibroblast cells were co-cultured with GC cells, and invasion, migration, epithelial-mesenchymal transformation (EMT) were determined. After blocking the expression of fibroblast CXCL12, cells were co-cultured with a GC cell line. Detection of GC cell line invasion, migration, EMT and CXCR4, Wnt5a and β-Catenin expression levels was performed. Primary CAFs and gastric normal fibroblasts were isolated and CXCL12 mRNA and protein expression were determined. In addition, a cohort of 285 GC cases was established, protein expression was evaluated immunohistochemically, and prognostic results were analyzed. Results: GC transcriptome analysis suggested that cytokine-cytokine receptor interaction and the Wnt signaling pathway in GC tissues were significantly up-regulated. scRNA analysis of advanced malignant GC samples showed that severe intestinal metaplasia (SIM) in GC specimens of different malignant grades had obvious fibroblast clusters compared to non-atrophic gastritis (NAG) and early gastric cancer (EGC). In the SIM group, fibroblast cluster, CXCL12, CXCR4, and Wnt5a were overexpressed. Co-culturing with fibroblast cells significantly increased the invasion, migration, and EMT of GC cells, and blocking CXCL12 in CAFs disturbed the expression of Wnt5a and β-catenin. In our cohort of GC patients, high CXCL12 expression in CAFs significantly correlated with histological grade (P = 0.012) and TNM stage (P = 0.014), as well as with poor overall survival (p = 0.0107). Conclusion: High expression of CXCL12 in CAFs in a GC microenvironment can affect the migration, invasion, and EMT of GC cells. Furthermore, it can cause poor prognosis in patients with GC.

Keywords: CAFs; CXCL12; gastric cancer; microenvironment; scRNA.