Effect of Regulatory and Angiogenic Cytokines RNA interference on Trophoblast Invasion.

DUBINSKY V; POEHLMANN T; GUTIERREZ G; GENTILE T; MARKERT U

Berlin, Alemania

Congreso; 5th European Congress of reproductive Immunology.; 2007

Sociedad Europea de Inmunología de la Reproducción

Trophoblast invasion is a temporal and local process which regulates implantation and oxygen arrival to the embryo trough the dialogue with spiral artery endothelium. These facts allow us to postulate that may exist a regulatory mechanism which ends trophoblast proliferation in order to facilitate cell migration to the decidual tissue and subsequent invasion. On the other hand, the end of implantation ruled by inflammatory cytokines and the begining of the angiogenic and anti-inflammatory placentation is essential for a normal pregnancy. We have previously demonstrated that placental IL-6 prevents recurrent abortion in CBA/J x DBA/2 murine model, whereas increases the levels of placental IL-6 and angiogenic cytokine VEGF. Regarding this result, we postulate that Hipoxia Inducible Factor- 1á (HIF-1á), a transcription factor for VEGF and IL-6, could be implicated in the regulation of IL-6 and VEGF expression in the feto-maternal interface. Considering also that Markert et all have recently demonstrated that IL-6 could be implicated in Stat3 regulation mechanism, we decided to investigate the effect of IL-6, VEGF and HIF-1á RNA interference in the trophoblast functionality in vitro. For this purpose we transfected JEG-3 cells with IL-6, VEGF and HIF-1á siRNA and scrambled siRNA as a control. Then we performed matrigel invasion and transwell migration assays and cell count by FACS. Cytokine knock-down effect was determined by ELISA and by western-blot for HIF-1á. We found a significant decrease in trophoblast migration when performed RNA interference for IL-6, VEGF and HIF-1á (p=0.029, 0.0072 and 0.031 respectively). Moreover, invasion was significantly diminished in JEG-3 cells transfected with IL-6 and HIF-1á siRNA (p=0.02 and 0.002 respectively) but not with VEGF siRNA (p=0.14). Taking these preliminary results into account we could infer that protective effect of IL-6 may be achieved by the regulation of embryo invasion which allows the modulation of developing feto-maternal circulation.