FKBP51 protein increase cell proliferation in HEK293 cells

BENDER T.; MAZAIRA G.I.; DE LEO S.A.; GALIGNIANA M.D.; CIUCCI S.; ERLEJMAN A.G

buenos aires

Congreso; LXIV Reunión anual de la Sociedad Argentina de Investigación Clínica (SAIC) en el marco de la Reunión Conjunta de Sociedades de Biociencias; 2019

Sociedad Argenitna de Investigación Clínica

The FK506-binding protein 51 (FKBP51) was first described as regulatory factor of steroid receptors. Currently, it is regarded that it also regulates diverse transcription factors (TFs). Previously, we demonstrated that FKBP51 modulates the transcriptional activity of NF-B, AP-1 and E2F. Because, these TFs participate dynamically in cell proliferation, our aim was to investigate the effect of FKBP51 on cell proliferation. HEK293 cells were transfected with pCI-Neo or pCI-Neo-hFKBP51 expression plasmids. After one month of selection the stable transfected clones were selected. HEK51 cells stably overexpress FKBP51 protein, in the same way, HEKpCI-Neo were selected containing the empty vector. Cell proliferation (viable cell count with Trypan Blue dye), proliferation index (flow cytometric analysis of cell division using dilution of CFSE) and cell cycle distribution (flow cytometry analysis of propidium iodide DNA stained) were evaluated. A Human Gene Expression Microarrays (SurePrint G3 Human GE v2 8x60K, Agilent) were performed for HEK51 vs. HEKpCI-Neo.After 96hs seeding, FKBP51 overexpression significantly duplicated cells proliferation of HEK51 relate to HEKwt (p=0,031). As well, FKBP51 duplicated the proliferation index from of 5.72 (HEKpCI-Neo) to 11.54 folds (HEK51). Interestingly, after 24h of starving HEK51 showed a 50,7 % of cells on S phase, while was 38,7% for HEKpCI-Neo. Gene expression microarray showed 272 differentially expressed transcripts (p≤0.001, BRB array tools-NIH). These genes grouped by biological function (Knowledge Data Base, Quiagen) showed that FKBP51 affected 81 molecules involved on cellular grown and proliferation.We demonstrate that FKBP51 increases proliferation of HEK293 cells. Therefore, this work suggests a relevant role for FKBP51 in cellular proliferation and deserves further thorough studies. Funding: UBACyT/CONICET. Acknowledgment: Drs. C.Robello and G.Greif, Institute Pasteur, Montevideo.