Trophoblast proliferation is inhibited by FKBP51

MAZAIRA,GISELA; FONTANA VANINA ANDREA; GALIGNIANA, MARIO; CAMISAY MARIA FERNANDA ; ERLEJMAN ALEJANDRA

buenos aires

Congreso; IFPA-SLIMP; 2019

IFPA-SLIMP

Understanding theregulation of proliferation of trophoblast is crucial to determine the etiologyof several obstetric disorders associated with the placenta. Previously, wedemonstrated, in trophoblast cells, that FK506 binding proteins (FKBPs)modulate the activity of transcription factors AP-1 and NF-kB, which regulate cell proliferation. Also, FKBP51and FKBP52 have been identified as regulators of steroid receptors. AlthoughFKBP51 and FKBP52 have high identity of sequence and structural similarity,they perform antagonist roles. While FKBP52 is a positive regulator, FKBP51functions as a negative regulator. Aim: to investigate the effect of FKBP51 on trophoblastproliferation. BeWo cell line was used as in vitro model of trophoblast cell. Cellswere transfected with pCI-Neo or pCI-Neo-hFKBP51 expression plasmids. After onemonth of selection with the antibiotic G418 (0.4 mg/ml), the stable transfectedclones were selected. BewoFKBP51 cells stably overexpress FKBP51 protein, inthe same way, BeWopCI-Neo were selected containing the empty vector. Cell proliferation (viable cell count withTrypan Blue dye) were evaluated for wildtype BeWo (BeWoWT), BewoFKBP51 and BeWopCI-Neo. As well, secretion ofinterleukin 6 (IL-6) (ELISA), proteolytic activity of the metalloproteinase matrix2 (MMP-2) (zymography), and colonies formation (counted after 10 days of seeding)were measured.After 72hs seeding, FKBP51 overexpression significantly inhibited cellsproliferation showing a relative cell proliferation of 31.9±8.5 BewoFKBP51 against 91.8±13.3 BeWopCI-Neo, p