miR-33a-3p regulates METTL3-mediated AREG stability and alters EMT to inhibit pancreatic cancer invasion and metastasis

Background: Recent studies have shown that amphoteric regulatory protein (AREG), a member of the epidermal growth factor (EGF) family, is expressed in many cancers and is an independent prognostic indicator for patients with pancreatic cancer, but whether AREG is regulated at the epigenetic level to promote the development of pancreatic cancer (PC) has not been elucidated. Methods: The expression level and role of AREG in PC cell tissues were investigated by GEPIA database, quantitative PCR (qPCR), Western blotting, CCK8 and Transwell assays. Methylated RNA immunoprecipitation (MeRIP) sequencing and RNA sequencing indicated that METTL3 regulates the m6A modification of AREG and affects its expression. RIP, MeRIP were used to explore its regulatory role and potential mechanisms. The association between AREG expression and clinical classification parameters was analyzed by Fisher’s exact test. In addition, we predicted that miR-33a-3p regulates METTL3 expression by miRDB and RNAinter. The intrinsic mechanisms and regulatory roles were explored using dual luciferase reporter assays and rescue experiments. Results: Our results support the notion that AREG is overexpressed in pancreatic cancer tissues and cell lines. Functionally, the deletion of AREG impedes pancreatic cancer (PC) cell proliferation, migration, and invasion. In addition, we identified and validated that methyltransferase-like 3 (METTL3) induced the m6A modification on AREG and facilitated the stability of AREG mRNA after sequencing. Additionally, we obtained experimental evidence that miR-33a-3p targets and inhibits METTL3 from taking action. Remediation experiments showed that miR-33a-3p inhibits PC progression through METTL3. Conclusions: this research reveals that miR-33a-3p inhibits m6A-induced stabilization of AREG by targeting METTL3, which plays a key role in the aggressive progression of PC. AREG could be a potential target for PC treatment.