Wu Quan, Shi Jiandang, Chen Linfeng, Wang Chun-Yu, Park Irwin, Lee Chung, Zhang Ju
Bioactive Materials Key Laboratory of Ministry of Education, Institute for Molecular Biology, Nankai University, Tianjin, PR China.
BJU Int. 2008 Feb;101(4):497-502. doi: 10.1111/j.1464-410X.2007.07340.x. Epub 2008 Jan 8.
To characterize a paracrine effect of prostatic epithelial cells in the presence or absence of oestradiol on the differentiation and proliferation of prostatic stromal cells.
Conditioned media (CM) collected from a prostatic epithelial cell line (BPH-1), which was pre-treated with different concentration of oestradiol, were added to cultures of primary prostatic stromal cells. The proliferation rates of stromal cells were determined using a tetrazolium assay. The mRNA level was analysed by real-time reverse transcription-polymerase chain reaction (RT-PCR), and the protein level of smooth muscle myosin heavy chain (SM-MHC), fibronectin and collagen IV were determined with Western blotting, enzyme- linked immunosorbent assay and radioimmunoassay, respectively. The expression of transforming growth factor beta1 (TGF beta 1) in the BPH-1 cell line was analysed.
The rate of proliferation of stromal cells increased when they were cultured with CM harvested from oestradiol-treated BPH-1 cells, but there was no remarkable change when they were cultured with CM from untreated cells. The level of smoothelin mRNA and SM-MHC protein increased after treatment with CM from BPH-1. The CM from BPH-1 with oestradiol stimulation was more effective in stimulating smoothelin mRNA and SM-MHC protein level. The protein level of collagen type IV, but not fibronectin, was up-regulated in the supernatants and cell extracts of CM-treated stromal cells. Oestradiol enhanced the expression and secretion of TGF beta 1 in BPH-1 cells. TGF beta 1-neutralizing antibody abrogated the effect of BPH-1 CM on the synthesis of collagen IV and SM-MHC in stromal cells.
These results suggest that oestradiol-stimulated proliferation and differentiation of prostatic stromal cells could be regulated by factors secreted from prostatic epithelial cells.
探讨在有或无雌二醇存在的情况下，前列腺上皮细胞对前列腺基质细胞分化和增殖的旁分泌作用。
将用不同浓度雌二醇预处理的前列腺上皮细胞系（BPH-1）收集的条件培养基（CM）添加到原代前列腺基质细胞培养物中。使用四氮唑盐测定法测定基质细胞的增殖率。通过实时逆转录-聚合酶链反应（RT-PCR）分析mRNA水平，分别用蛋白质印迹法、酶联免疫吸附测定法和放射免疫测定法测定平滑肌肌球蛋白重链（SM-MHC）、纤连蛋白和IV型胶原的蛋白质水平。分析BPH-1细胞系中转化生长因子β1（TGFβ1）的表达。
与从经雌二醇处理的BPH-1细胞收获的CM一起培养时，基质细胞的增殖率增加，但与未经处理的细胞的CM一起培养时无明显变化。用BPH-1的CM处理后，平滑肌肌动蛋白mRNA和SM-MHC蛋白水平升高。经雌二醇刺激的BPH-1的CM在刺激平滑肌肌动蛋白mRNA和SM-MHC蛋白水平方面更有效。在CM处理的基质细胞的上清液和细胞提取物中，IV型胶原的蛋白质水平上调，而纤连蛋白则未上调。雌二醇增强了BPH-1细胞中TGFβ1的表达和分泌。TGFβ1中和抗体消除了BPH-1 CM对基质细胞中IV型胶原和SM-MHC合成的影响。
这些结果表明，前列腺上皮细胞分泌的因子可调节雌二醇刺激的前列腺基质细胞的增殖和分化。
