KLF6: A POSSIBLE NOVEL REGULATOR OF VILLOUS TROPHOBLAST FUSION

RACCA, ANA C.; RIDANO, MAGALÍ EVELIN; CAMOLOTTO, SOLEDAD; GENTI-RAIMONDI, SUSANA; PANZETTA-DUTARI, GRACIELA

Toronton

Congreso; 36th Annual Eastern Canadian Perinatal Investigators Meeting; 2012

Eastern Canadian Perinatal Investigators

OBJECTIVE The aim of this work is to elucidate Kruppel Like Factor 6 (KLF6) putative role in trophoblast cells during placentation. KLF6 is a transcription factor involved in differentiation, proliferation, cell cycle control and tumorigenesis. It is highly expressed in placenta and it has been reported that klf6 -/- knockout mice die by embryonic day 12.5, characterized by impaired placental development (Matsumoto et al 2006). With this background, we hypothesize that KLF6 could be involved in placenta development. METHODS To address this objective we used siRNA and overexpression of KLF6 experiments in primary villous cytotrophobast and JEG-3 cells, and analysed morphological fusion of primary CTB by desmoplakin staining and, by real time RT-PCR measured mRNA levels of genes that encode for proteins involved in the fusion process such as: Conexin-43, Syncitin-1 and 2 and Tight junction protein ZO-1 in transfected cells compared with controls. RESULTS Primary trophoblasts cells are disable to fuse when KLF6 is silenced, and in contrast, when KLF6 is overexpressed Conexin-43 is 3.012, Syn-1 3.968, Syn-2 8.257 and ZO-1 5.075 times higher than the control, but the rate of increase is highly dependent on the placenta. Also, in the JEG-3 model, cells that do not fuse spontaneously, the sustained overexpression of KLF6 increase Conexin-43, Syn-1, Syn-2 and ZO-1 mRNA levels at 2.620, 2.715, 4.029 and 2.017 times respectively. CONCLUSIONS We have previously demonstrated that KLF6 is strictly regulated during villous trophoblast differentiation, increasing fast and transiently at the beginning of the process and presenting a second wave of expression by the end of the syncytium formation. Present results allow us to suggest KLF6 as a possible novel regulator of villous trophoblast fusion.