CBX2 promoted oral squamous cell carcinoma via increasing CEP55/NF-κB/METTL3/SHP2 signaling induced metastasis/proliferation and angiogenesis

To investigate the mechanism by which CBX2 promotes oral squamous cell carcinoma (OSCC) by increasing CEP55/NF-κB/METTL3/SHP2 signaling, leading to metastasis, proliferation, and angiogenesis. We employed a comprehensive research approach. Through bioinformatics analysis (GSE153918 dataset), we identified differentially expressed genes (DEGs) and enriched pathways associated with OSCC and CBX2. Immunohistochemistry (IHC) was used to detect CBX2 expression in both tumor and peritumoral tissues from OSCC patients. Twenty-four BALB/c nude mice were divided into three groups and infected with or without CBX2-KD and/or METTL3-OE lentiviruses. Tumor volume and weight changes were observed in nude mice. IHC was performed to observe the protein expression of CBX2 and METTL3. The oral squamous cell carcinoma cell line SCC-25 was divided into six groups and subjected to CBX2 knockdown, NF-κB activation, METTL3 inhibition, METTL3 overexpression, and SHP2 inhibition. Western blot was used to observe the protein expression of CBX2, CEP55, p-NF-κB, METTL3, TGFβ1, p-SHP2, p-PI3K, Slug and Snail. Using Co-Immunoprecipitation (Co-IP) technology, we analyzed the interactions between endogenous CBX2 and CEP55, as well as the interactions between METTL3 and SHP2, and between METTL3 and TGFβ1, in cells. Wound healing, Transwell, and colony formation assays were conducted to observe the migration, invasion, and proliferation of tumor cells. Western blot was used to analyze the protein expression of VEGFA and H1F1α, and tube formation assays was performed to observe the effects on angiogenesis. Bioinformatic analysis revealed the critical role of the CBX2-NF-κB-METTL3-SHP2 pathway in oral squamous cell carcinoma. IHC results showed that CBX2 expression was lower in peritumoral tissues and higher in OSCC tissues. Tumor formation and IHC results in nude mice showed that CBX2 knockdown inhibited tumor growth, and METTL3 overexpression reversed this effect. Western blot results indicated that CBX2 knockdown, NF-κB activation, METTL3 inhibition, METTL3 overexpression, and SHP2 inhibition all upregulated or downregulated downstream signaling molecules. Co-IP experiments indicated that endogenous CBX2 and CEP55 physically interacted in SCC-25 cells, and METTL3 specifically interacted with SHP2 and TGFβ1. Wound healing, Transwell, and colony formation assays showed that CBX2 knockdown, NF-κB activation, METTL3 inhibition, METTL3 overexpression, and SHP2 inhibition all inhibited or promoted tumor cell migration, invasion, and proliferation. Western blot and tube formation assay results showed that CBX2 knockdown, NF-κB activation, METTL3 inhibition, METTL3 overexpression, and SHP2 inhibition all inhibited or promoted angiogenesis. CBX2 promoted oral squamous cell carcinoma via increasing CEP55/NF-κB/METTL3/SHP2 signaling, leading to metastasis, proliferation, and angiogenesis.