Trophoblast proliferation is inhibited by FKBP51

FONTANA, V; CAMISAY, MF; MAZAIRA, G; GALIGNIANA, M; ERLEJAMN, A

Buenos Aires

Congreso; Federation of Placenta Association (IFPA); 2019

Federation of Placenta Association (IFPA)

Understanding the regulation of proliferation of trophoblast is crucial to determine the etiology of several obstetric disorders associated with the placenta. Previously, we demonstrated, in trophoblast cells, that FK506 binding proteins (FKBPs) modulate the activity of transcription factors AP-1 and NF-B, which participate in proliferation regulation. Also, FKBP51 and FKBP52 have been identified as regulators of steroid receptors. Although FKBP51 and FKBP52 have high identity of sequence and structural similarity, they perform antagonist roles. While FKBP52 is a positive regulator, FKBP51 functions as a negative regulator.To investigate the effect of FKBP51 on trophoblast proliferation and invasion BeWo cell line was used as in vitro model of trophoblast cell. Cells were transfected with pCI-Neo or pCI-Neo-hFKBP51 expression plasmids. After one month of selection with the antibiotic G418 (0.4 mg/ml), the stable transfected clones were selected. BewoFKBP51 cells stably overexpress FKBP51 protein, in the same way, BeWopCI-Neo were selected containing the empty vector. Cell proliferation (viable cell count with Trypan Blue dye) were evaluated for wild type BeWo (BeWoWT), BewoFKBP51 and BeWopCI-Neo. As well, secretion of interleukin 6 (IL-6) (ELISA), proteolytic activity of the metalloproteinase matrix 2 (MMP-2) (zymography), and colonies formation (counted after 10 days of seeding) were measured.After 72hs seeding, FKBP51 overexpression significantly inhibited cells proliferation showing a relative cell proliferation of 31.9±8.5 BewoFKBP51 against 91.8±13.3 BeWopCI-Neo, p