WTAP stabilizes MMP12 expression to promote the malignant phenotypes of esophageal cancer cells

WTAP通过稳定MMP12表达促进食管癌细胞的恶性表型。

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Abstract

BACKGROUND: Esophageal cancer (EC) is a highly aggressive malignancy with a poor prognosis, largely due to its invasive nature and metabolic reprogramming. Matrix metalloproteinase 12 (MMP12) has been implicated in various cancers, but its specific regulatory mechanisms and functional impact on EC remain unclear. This study aimed to investigate the role of MMP12 in EC progression and its underlying regulatory mechanisms. METHODS: The GSE161533 dataset was analyzed to identify differentially expressed genes (DEGs) between EC and normal tissues, with a focus on genes linked to apoptosis, proliferation, and glycolysis via the GeneCards database. Key signature genes were further screened using lasso regression, support vector machine (SVM), and random forest (RF) algorithms. Gene expression was validated by quantitative real-time polymerase chain reaction and Western blotting. Cell migration and invasion were analyzed by transwell assays. Cell proliferation was analyzed by 5-Ethynyl-2'-deoxyuridine assay. Cell apoptosis was assessed by flow cytometry. Glucose consumption, lactate production, and ATP levels were analyzed by commercial kits. Flow cytometry was used to quantify the number of clusters of differentiation 68 (CD68)-positive cells and CD206-positive macrophages. The interaction between MMP12 and WT1 associated protein (WTAP) was examined using methylated RNA immunoprecipitation (MeRIP) and RNA immunoprecipitation (RIP) assays. RESULTS: MMP12 was identified as a key gene in EC tissues, and its expression was significantly upregulated in EC tissues and cells in comparison with normal samples. Knockdown of MMP12 suppressed migration, invasion, proliferation, and glycolysis while promoting apoptosis in KYSE150 and TE-10 cells. Additionally, MMP12 silencing inhibited M2 macrophage polarization. Mechanistically, WTAP stabilized MMP12 expression in an m6A-dependent manner. Further, silencing WTAP suppressed the malignant phenotypes of KYSE150 and TE-10 cells by downregulating MMP12 expression. CONCLUSION: WTAP stabilized MMP12 expression via m6A modification, thereby enhancing the malignant phenotypes of EC cells. Clinically, targeting the WTAP-MMP12 axis could represent a promising therapeutic strategy to inhibit EC progression.

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