Estradiol stimulates cell proliferation in primary cultures of human renal tubular epitelial cells

DAIANA S SANCHEZ; LILIAN K FISCHER SIGEL; ELISABET M ODDO; CLAUDIA SILBERSTEIN; PABLO J AZURMENDI; FERNANDO R IBARRA

Mar del Plata

Congreso; LXIII Reunión Anual de la Sociedad Argentina de Investigación Clínica (SAIC), de la Sociedad Argentina de Fisiología (SAFIS) y LXVI Sociedad Argentina de Inmunología (SAI).; 2018

Sociedad Argentina de Investigación Clínica

We have previously demonstrated that 17β-Estradiol (E2) stimulatescell proliferation through classic estrogen receptors (ER) and the Gprotein-coupled estrogen receptor 1 (GPER-1), in primary cultures ofhuman cortical renal tubular epithelial cells (HRTEC). The aim of thepresent work was to study the effects of E2 on cell proliferation andto evaluate the estrogen receptor and intracellular signals involvedin this mechanism in HRTEC. Primary cultures were developed fromnephrectomies performed in pediatric patients at the Hospital NacionalProf. A. Posadas. HRTEC were treated with E2 (10 nM for 24h) with or without classic ER antagonist, ICI 182,780, and GPER-1agonist and antagonist, G-1 and G-15, respectively. Cell proliferationrate was measured by incorporation of 5-bromo-2-deoxyuridine(BrdU) in nuclei of HRTEC primary cultures, and by counting the cellnumber at 24 and 48 h. ERalpha, ERbeta, β-catenin, and cyclin D1expression and localization were assayed by western blot analysisand immunofluorescence, respectively. Treatment with any of bothE2 and G-1 (10 nM, 24 h) stimulated significantly the BrdU uptake inHRTEC primary cultures, and increased the number of cells at 48 h.Co-incubation of HRTEC with E2 and G-1 increased BrdU uptake asoccurred by E2 alone. The treatment of HRTEC with E2 significantlyincreased CyD1 and β-catenin protein expression, and increasedthe number of cells that translocated ERalpha and β-catenin intotheir nucleus. However, E2 did not modified ERbeta expressionand localization, compared to control HRTEC. Both G-15 and ICI182,780 totally abrogated E2- stimulation of BrdU uptake, and β-cateninexpression and translocation to the nucleus in HRTEC. GPER-1 expression and localization was not modified by E2 treatment. Inconclusion, E2 regulates HRTEC proliferation by activating both estrogensreceptors, ERα and GPER-1, possibly by stimulating β-cateninpathway, in normal human renal tubular epithelium.