Hirschsprung's disease: m6A methylase VIRMA suppresses cell migration and proliferation by regulating GSK3

BackgroundN6-methyladenosine (m6A) is the most abundant mRNA modification in mammals, participating in various biological processes. VIRMA is a key methyltransferase involved in m6A modification. However, the role of VIRMA in Hirschsprung's disease (HSCR) remains unclear. This study aims to investigate the function of VIRMA in HSCR and identify its corresponding regulatory mechanisms.MethodsThe expression of VIRMA and GSK3 beta in colon tissues of HSCR was examined using RT-qPCR, Western blot, and Immunohistochemistry. Immunofluorescence detected localization of VIRMA and GSK3 beta. Cell proliferation was measured by CCK8 and EdU assays, and cell migration was evaluated via cell migration and wound healing assays. The stability of GSK3 beta mRNA was assessed using the actinomycin D assay and the overall level of m6A in cells was assessed by colorimetric assay.ResultsVIRMA was significantly downregulated in narrow-segment colon tissue. Silencing of VIRMA inhibited cell proliferation and migration. VIRMA can inhibit the degradation of GSK3 beta mRNA and increase the expression of GSK3 beta. GSK3 beta was significantly upregulated in narrow-segment colon tissues. Accordingly, our findings showed that GSK3 beta mediated the VIRMA-driven cell migration and proliferation.ConclusionVIRMA can inhibit cell migration and proliferation by upregulating the expression of GSK3 beta, contributing to the onset of HSCR.ImpactThe expressions of VIRMA were significantly reduced in HSCR, while GSK3 beta expression was increased in HSCR, and can be used as a molecular marker.VIRMA overexpression promoted the proliferation and migration of SH-SY5Y and HEK-293T cells.VIRMA can inhibit the degradation of GSK3 beta mRNA and increase the expression of GSK3 beta.