NF-kappaB decoy oligonucleotides suppress RANTES expression and monocyte chemotactic activity via NF-kappaB inactivation in stromal cells of ectopic endometrium.
BACKGROUND
The nuclear factor kappa B (NF-kappaB) 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.


OBJECTIVE
The objective of this study was to investigate the effect of NF-kappaB decoy oligonucleotides (ODNs) on NF-kappaB activation, RANTES expression, and monocyte chemotactic activity in ectopic endometrial stromal cells in vitro.


METHODS
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)-1beta. Four hours after transfection of NF-kappaB decoy ODNs, 10 ng/ml IL-1beta was added to induce the ectopic endometrial stromal cells to secrete RANTES. The NF-kappaB activation, RANTES expression, and monocyte chemotactic activity in ectopic endometrial stromal cells were respectively evaluated by electrophoretic mobility shift assay, ELISA, and Boyden chambers.


RESULTS
IL-1beta induced significantly higher levels (P < 0.05) of RANTES expression in a time-dependent manner in ectopic endometrial stromal cells compared with IL-1beta-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-kappaB decoy ODNs dramatically decreased (P < 0.05) the NF-kappaB activation, RANTES expression, and monocyte chemotactic activity in IL-1beta-induced ectopic endometrial stromal cells.


CONCLUSIONS
NF-kappaB decoy ODNs may exert anti-inflammatory effects in ectopic endometrial stromal cells via the suppression of NF-kappaB activation, RANTES expression, and monocyte chemotactic activity.
Xiu-li
Wang
W
Department of Obstetrics and Gynecology, The First Affiliated Hospital of Nanjing Medical University, 368 North-Jiangdong Road, Nanjing, China. xiuli_2266@163.com
Su-ping
Han
H
Hui-hua
Dai
D
Zhi-xue
You
Y
Shi-long
Fu
F
Pin-hong
Lu
L
D002464:Cell Migration Inhibition; D018946:Chemokine CCL5 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002828:Choristoma / Q000378:metabolism / Q000473:pathology; D004715:Endometriosis / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D004717:Endometrium / Q000378:metabolism* / Q000473:pathology; D005260:Female; D006801:Humans; D009000:Monocytes / Q000276:immunology / Q000378:metabolism*; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D009841:Oligonucleotides; D000292:Pelvic Inflammatory Disease / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D011247:Pregnancy; D017154:Stromal Cells / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D052416:Targeted Gene Repair; D014162:Transfection
D018946:Chemokine CCL5; D016328:NF-kappa B; C468119:NF-kappaB decoy; D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides