Areca nut‐induced AREG promote oral epithelial cell proliferation, migration, and EMT

Abstract Objective To analyze the biological effect and mechanism of areca nut extract (ANE) on human oral keratinocyte (HOK) cells. Materials and Methods The effect of gradient concentration of ANE on the proliferation activity of HOK cells was analyzed by cell counting kit‐8 (CCK‐8) assays. The differentially expressed genes between the ANE group and control group HOK cells were analyzed by second‐generation transcriptome sequencing. Real‐time PCR and western blot were, respectively, used to analyze the expression of AREG gene and protein in HOK cells. After AREG gene overexpression or knockdown, the proliferation, migration, and expression of proteins related to epithelial–mesenchymal transformation (EMT), MAPK signal pathway in HOK cells were, respectively, detected by CCK‐8, wound healing, transwell, and western blot assays. Results ANE (500 μg/mL) promoted the proliferation and migration of HOK cells, ANE (2 mg/mL) promoted the EMT of HOK cells, and ANE (50 mg/mL) inhibited the proliferation of HOK cells. AREG knockdown inhibited ANE‐induced proliferation and migration of HOK cells, while AREG overexpression promoted the proliferation and migration of HOK cells. Western blot assay showed that ANE activated MAPK signal pathway by upregulating AREG protein in HOK cells. Conclusions ANE promoted HOK cell proliferation, migration, and EMT by mediating AREG–MAPK signaling pathway.