Role and mechanism of the CXCL12/CXCR4 axis in pediatric neurofibromatosis type Ⅰ
Lu Yuanfang , Guo Xiaolan , Yang Aiyun, et al
(1. Beijing Key Laboratory of Environment and Viral Oncology, Faculty of Environment and Life Sciences, Beijing University of Technology, Beijing 100124, China; 2. Beijing Key Laboratory of Child Development and Nutriomics, Translational Medicine Laboratory, Capital Institute of Pediatrics, Beijing 100020, China; 3. Department of Neurology, Capital Center for Children’s Health, Capital Medical University, Institute of Basic Medicine, Chinese Academy of Medical Sciences, Beijing 100020, China; 4. School of Basic Medicine, Peking Union Medical College, Institute of Basic Medicine, Chinese Academy of Medical Sciences, Beijing 100005, China; 5. Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200003, China)
Abstract: 【Abstract】 Objective To investigate the role and mechanism of the C-X-C chemokine ligand 12/C-X-C chemokine receptor 4 (CXCL12/CXCR4) axis in neurofibromatosis type I (NF1) in children.Method The GSE241224 dataset was obtained from the Gene Expression Omnibus (GEO) database, and compared the expression of CXCL12 between malignant peripheral nerve sheath tumor (MPNST) and benign neurofibromas in children with NF1. Weighted gene co-expression network analysis (WGCNA) was conducted on the top 5 000 high-variance genes to identify the module most significantly correlated with the CXCL12 gene, followed by gene ontology (GO) functional enrichment analysis. The expression profile of the CXCR4 gene in tumor and normal tissues was analyzed using the gene expression profiling interactive analysis (GEPIA) database. Human Schwann cells (ipn02.3 2λ), patient-derived plexiform neurofibroma (PNF) cell line (WZJ), and MPNST cell lines (S462 and sNF96.2) were utilized, CXCR4 expression was detected by flow cytometry and Western blotting, and its effect in cell migration was investigated using the CXCR4-specific inhibitor AMD3100 through a cell scratch assay. Statistical analysis was performed by analysis of variance and Dunnett's t-test, and the bioinformatics data were analyzed using R language. Result The expression of CXCL12 in MPNST was significantly higher than in the benign neurofibroma group (P=0.041 1), with 1 630 genes upregulated and 877 genes downregulated. The expression levels of CXCL12 showed positive correlations with genes such as microRNA MIR3687 (r=0.57, P=3.72×10-8), E3 ubiquitin ligase gene RNF185 (r=0.58, P=1.53×10-8), Zinc-binding dehydrogenase gene ZADH2 (r=-0.58, P=1.58×10-8), sialyltransferase gene ST8SIA3 (r=0.59, P=1.43×10-8), and Zinc finger protein gene ZNF791(r=0.50, P=6.83×10-9). The expression levels of CXCL12 showed significant negative correlations with the microtubule-severing enzyme gene KATNAL1 (r=-0.58, P=2.56×10-6), migration and invasion inhibitory protein gene MIIP (r=-0.52, P=1.16×10-6), and deubiquitinase gene USP42 (r=-0.52, P=9.40×10-7). WGCNA analysis revealed that the blue module had the highest correlation with CXCL12 (r=-0.46, P<0.001), with its genes enriched in biological functions such as regulation of the cell cycle, positive regulation of DNA metabolic processes, protein transport from the Golgi to the plasma membrane, cellular component disassembly, and organelle localization. The three genes exhibiting the strongest correlations were autophagy-related 4C cysteine peptidase (ATG4C), pleckstrin homology domain containing, family M with Rundomia member M2 (PLEKHM2), and Zinc finger protein 140 (ZNF140). In NF1-related neural tumors, the expression of CXCR4 was significantly higher than in normal tissues (P<0.05). The results of flow cytometry showed that the positivity rates of CXCR4 on the cell membrane surfaces of ipn02.3 2λ, WZJ, S462, and sNF96.2 were 2.52%, 0.18%, 2.03%, and 0.57%, respectively, while the intracellular positivity rates were 89.7%, 79.3%,94.3%, and 96.4%, respectively. The results of Western blotting confirmed that CXCR4 was highly expressed in MPNST cells. Treatment with the CXCR4-specific inhibitor AMD3100 significantly inhibited the migration of MPNST cells [(38.05±10.58) % ]compared to ipn02.3 2λcells [(60.30±2.43)% ](P=0.026). ConclusionThe CXCL12/CXCR4 axis is highly expressed in pediatric NF1, particularly in MPNST, which promoted the migration of malignant cells. It may also influence gene expression in NF1 cells through genes such as MIR3687, RNF185, ZADH2, ST8SIA3, ZNF791, KATNAL1, MIIP, USP42, ATG4C, PLEKHM2 and ZNF140, thereby affects the development and progression of MPNST
路媛芳 郭晓兰 杨爱云 等. CXCL12/CXCR4 轴在儿童期神经纤维瘤I 型中的作用及机制[J]. 发育医学电子杂志, 2025, 13(4): 241-251.
Lu Yuanfang , Guo Xiaolan , Yang Aiyun, et al. Role and mechanism of the CXCL12/CXCR4 axis in pediatric neurofibromatosis type Ⅰ. Journal of Developmental Medicine(Electronic Version), 2025, 13(4): 241-251.
2025, Vol. 13 
