title: Destabilization of TNF-α mRNA by Rapamycin.
abstract: Stimulation of mast cells through the high affinity IgE receptor (FcεRI) induces degranulation, lipid mediator release, and cytokine secretion leading to allergic reactions. Although various signaling pathways have been characterized to be involved in the FcεRI-mediated responses, little is known about the precious mechanism for the expression of tumor necrosis factor-α (TNF-α) in mast cells. Here, we report that rapamycin, a specific inhibitor of mammalian target of rapamycin (mTOR), reduces the expression of TNF-α in rat basophilic leukemia (RBL-2H3) cells. IgE or specific antigen stimulation of RBL-2H3 cells increases the expression of TNF-α and activates various signaling molecules including S6K1, Akt and p38 MAPK. Rapamycin specifically inhibits antigen-induced TNF-α mRNA level, while other kinase inhibitors have no effect on TNF-α mRNA level. These data indicate that mTOR signaling pathway is the main regulation mechanism for antigen-induced TNF-α expression. TNF-α mRNA stability analysis using reporter construct containing TNF-α adenylate/uridylate-rich elements (AREs) shows that rapamycin destabilizes TNF-α mRNA via regulating the AU-rich element of TNF-α mRNA. The antigen-induced activation of S6K1 is inhibited by specific kinase inhibitors including mTOR, PI3K, PKC and Ca(2+)chelator inhibitor, while TNF-α mRNA level is reduced only by rapamycin treatment. These data suggest that the effects of rapamycin on the expression of TNF-α mRNA are not mediated by S6K1 but regulated by mTOR. Taken together, our results reveal that mTOR signaling pathway is a novel regulation mechanism for antigen-induced TNF-α expression in RBL-2H3 cells.
authors.affiliation: Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Jong-Woo
authors.initials: JW
authors.lastname: Park
authors.forename: Ye Ji
authors.initials: YJ
authors.lastname: Jeon
authors.forename: Jae Cheol
authors.initials: JC
authors.lastname: Lee
authors.forename: So Ra
authors.initials: SR
authors.lastname: Ahn
authors.forename: Shin Won
authors.initials: SW
authors.lastname: Ha
authors.forename: So Young
authors.initials: SY
authors.lastname: Bang
authors.forename: Eun Kyung
authors.initials: EK
authors.lastname: Park
authors.forename: Sang Ah
authors.initials: SA
authors.lastname: Yi
authors.forename: Min Gyu
authors.initials: MG
authors.lastname: Lee
authors.forename: Jeung-Whan
authors.initials: JW
authors.lastname: Han
keywords: Mast cells; Rapamycin; S6K1; Stability; mRNA; mTOR