NF-κB Decoy Oligonucleotides Suppress RANTES Expression and Monocyte Chemotactic Activity via NF-κB Inactivation in Stromal Cells of Ectopic Endometrium

The nuclear factor kappa B (NF-kappa B) pathway is a critical mediator of regulated on activation, normal T cell expressed and secreted (RANTES) gene regulation and therefore represents a potential target for therapy of endometriosis-associated symptoms. The objective of this study was to investigate the effect of NF-kappa B decoy oligonucleotides (ODNs) on NF-kappa B activation, RANTES expression, and monocyte chemotactic activity in ectopic endometrial stromal cells in vitro. A specific sandwich enzyme-linked immunosorbent assay (ELISA) was used to quantify RANTES expression in ectopic and normal endometrial stromal cells stimulated by interleukin (IL)-1 beta. Four hours after transfection of NF-kappa B decoy ODNs, 10 ng/ml IL-1 beta was added to induce the ectopic endometrial stromal cells to secrete RANTES. The NF-kappa B activation, RANTES expression, and monocyte chemotactic activity in ectopic endometrial stromal cells were respectively evaluated by electrophoretic mobility shift assay, ELISA, and Boyden chambers. IL-1 beta induced significantly higher levels (P < 0.05) of RANTES expression in a time-dependent manner in ectopic endometrial stromal cells compared with IL-1 beta-untreated ectopic and normal endometrial stromal cells. The RANTES accounts for the majority (68%) of the monocyte chemotactic activity in conditioned media of ectopic endometrial stromal cells. In vitro transfection of NF-kappa B decoy ODNs dramatically decreased (P < 0.05) the NF-kappa B activation, RANTES expression, and monocyte chemotactic activity in IL-1 beta-induced ectopic endometrial stromal cells. NF-kappa B decoy ODNs may exert anti-inflammatory effects in ectopic endometrial stromal cells via the suppression of NF-kappa B activation, RANTES expression, and monocyte chemotactic activity.