Cox-2-derived PGE₂ induces Id1-dependent radiation resistance and self-renewal in experimental glioblastoma

Background In glioblastoma (GBM), Id1 serves as a functional marker for self-renewing cancer stem-like cells. We investigated the mechanism by which cyclooxygenase-2 (Cox-2)-derived prostaglandin E₂ (PGE₂) induces Id1 and increases GBM self-renewal and radiation resistance. Methods Mouse and human GBM cells were stimulated with dimethyl-PGE₂ (dmPGE₂), a stabilized form of PGE₂, to test for Id1 induction. To elucidate the signal transduction pathway governing the increase in Id1, a combination of short interfering RNA knockdown and small molecule inhibitors and activators of PGE₂ signaling were used. Western blotting, quantitative real-time (qRT)-PCR, and chromatin immunoprecipitation assays were employed. Sphere formation and radiation resistance were measured in cultured primary cells. Immunohistochemical analyses were carried out to evaluate the Cox-2-Id1 axis in experimental GBM. Results In GBM cells, dmPGE₂ stimulates the EP4 receptor leading to activation of ERK1/2 MAPK. This leads, in turn, to upregulation of the early growth response1 (Egr1) transcription factor and enhanced Id1 expression. Activation of this pathway increases self-renewal capacity and resistance to radiation-induced DNA damage, which are dependent on Id1. Conclusions In GBM, Cox-2-derived PGE₂ induces Id1 via EP4-dependent activation of MAPK signaling and the Egr1 transcription factor. PGE₂-mediated induction of Id1 is required for optimal tumor cell self-renewal and radiation resistance. Collectively, these findings identify Id1 as a key mediator of PGE₂-dependent modulation of radiation response and lend insight into the mechanisms underlying radiation resistance in GBM patients.