Estradiol stimulated proliferation and differentiation of prostatic stromal cells through regulation of BPH-1 paracrine / 中国病理生理杂志

ABSTRACT

AIM:To characterize the effect of estradiol on proliferation,differentiation and extracellular matrix(ECM) accumulation in stromal cells through regulation of BPH-1 paracrine.METHODS:BPH-1 cells were stimulated with different concentrations of estradiol.Conditioned media(CM) were harvested and their effects on stromal cell cultures were tested.Cell proliferation was determined by MTT assay.mRNA of smoothelin,fibronectin,collagen Ⅳ and transforming growth factor ?1(TGF-?1) were analyzed by real-time RT-PCR.Western blotting was used to determine smooth muscle myosin heavy chain(SMMHC).ELISA and radioimmunoassay were respectively used to measure fibronectin,TGF-?1 and collagen Ⅳ protein expressions.RESULTS:Estrodiol stimulated the expression and secretion of TGF-?1 in BPH-1 cells.The proliferation of stromal cells increased when they were cultured with CM harvested from estrogen treated BPH-1 cells.The mRNA levels of collagen Ⅳ and smoothelin increased in stromal cells treated with CM from BPH-1 cells.The results of radioimmunoassay also showed that the collagen Ⅳ protein level up-regulated in the supernatants and cell extracts of CM-treated stromal cells.A neutralizing antibody to TGF-?1 inhibited the stimulation of collagen Ⅳ and SMMHC by BPH-1 CM.The expression of fibronectin was only marginally changed in stromal cells cultured in the presence of BPH-1 CM.CONCLUSION:The BPH-1 cells increase ECM accumulation and differentiation of stromal cells through TGF-?1.Estradiol stimulate differentiation of stromal cells by induction of TGF-?1 expression.Estradiol stimulate proliferation by influencing the factors secreted from prostatic epithelial cells.