The role and regulatory mechanism of IL‐1β on the methylation of the NF2 gene in benign meningiomas and leptomeninges. Issue 12 (3rd February 2016) - Wang, Baofeng
Abstract : Methylation of the neurofibromatosis type 2 (NF2) gene in low‐grade meningioma (WHO grade I) has crucial roles in tumorigenesis and development. Meningioma formation might also occur in the setting of an inflammatory microenvironment. However, the association between inflammation and the methylation of NF2 remains unclear. The present study investigates the role and regulatory mechanism of IL‐1β, one of the most important pro‐inflammatory cytokines, in the methylation of NF2 in benign meningioma. Three primary low‐grade meningioma cells and leptomeningeal cells were cultured. CCK‐8 and BrdU assays demonstrated that proliferation of meningioma/leptomeningeal cells treated with IL‐1β occurred in a dose‐ and time‐dependent manner. Methylation‐specific PCR verified that IL‐1β induced methylation of the NF2 promoter and decreased NF2/merlin expression in meningioma/leptomeningeal cells. Real‐time PCR, western blotting, and immunofluorescence showed that IL‐1β up‐regulated DNMT1 in meningioma cells and DNMT1/3b in leptomeningeal cells but did not up‐regulate DNMT3a. After co‐treatment with the DNMT inhibitor 5‐Aza‐2′‐deoxycytidine and DNMT siRNA, methylation of NF2 induced by IL‐1β was attenuated and merlin expression was restored. Furthermore, we showed that DNMT1 in meningiomas and DNMT1/3b in leptomeninges were regulated via activation of the MAPK (p38, ERK, JNK) and NF‐κB pathways. These results suggest that IL‐1β induces methylation of NF2 by up‐regulating DNMT1 in benign meningioma cells and DNMT1/3b in leptomeningeal cells via MAPK and NF‐κB pathways. Therefore, NF2 methylation is a linker between IL‐1β and tumor development, and DNMTs might be potential therapeutic targets in meningioma for regulating NF2 and inhibiting tumor development. © 2016 Wiley Periodicals, Inc.