Sherwood E R, Fong C J, Lee C, Kozlowski J M
Department of Urology, Northwestern University Medical School, Chicago, Illinois 60611.
Endocrinology. 1992 May;130(5):2955-63. doi: 10.1210/endo.130.5.1374018.
Studies were undertaken, using isolated prostatic epithelial and stromal cells, to evaluate the role of basic fibroblast growth factor (bFGF) in the regulation of benign prostatic growth. bFGF was detected in lysates, but not the conditioned media, of cultured prostatic epithelial and stromal cells by Western immunoblotting and immunoprecipitation of metabolically labeled proteins. Immunofluorescence analysis of benign human prostate localized the majority of bFGF to the prostatic stroma. In addition, bFGF was a potent stimulator of stromal cell proliferation in vitro, but was not mitogenic to cultured epithelial cells. Further studies demonstrated bFGF receptors (Kd = 258 pM; 61,400 receptors/cell) on stromal cells, but not epithelial cells. Epithelial cell-conditioned medium was mitogenic for stromal cells, suggesting the presence of paracrine interactions. However, bFGF does not appear to be the mediator of this interaction, since the mitogenic effect of epithelial cell-conditioned medium on stromal cells was not significantly reduced by the addition of anti-bFGF. Additional studies showed that concentrated stromal cell-conditioned medium was not mitogenic to cultured stromal cells under serum-free defined conditions, indicating the lack of an external autocine mechanism. These studies demonstrate that bFGF is actively synthesized by isolated prostatic epithelial and stromal cells, but is largely not secreted. Prostatic stroma, but not epithelia, are responsive to the mitogenic effect of bFGF in vitro. However, because of the limited secretion of bFGF by prostatic cells, the mechanism(s) of bFGF-mediated regulation of stromal growth remains unclear.
研究采用分离的前列腺上皮细胞和基质细胞,以评估碱性成纤维细胞生长因子(bFGF)在良性前列腺生长调节中的作用。通过对代谢标记蛋白进行Western免疫印迹和免疫沉淀,在培养的前列腺上皮细胞和基质细胞的裂解物中检测到bFGF,但在条件培养基中未检测到。对良性人前列腺的免疫荧光分析将大部分bFGF定位于前列腺基质。此外,bFGF在体外是基质细胞增殖的有效刺激剂,但对培养的上皮细胞没有促有丝分裂作用。进一步研究表明,基质细胞上存在bFGF受体(解离常数=258 pM;61400个受体/细胞),而上皮细胞上没有。上皮细胞条件培养基对基质细胞有促有丝分裂作用,提示存在旁分泌相互作用。然而,bFGF似乎不是这种相互作用的介质,因为添加抗bFGF并没有显著降低上皮细胞条件培养基对基质细胞的促有丝分裂作用。额外的研究表明,在无血清限定条件下,浓缩的基质细胞条件培养基对培养的基质细胞没有促有丝分裂作用,表明缺乏外部自分泌机制。这些研究表明,分离的前列腺上皮细胞和基质细胞可主动合成bFGF,但大部分不分泌。前列腺基质而非上皮对bFGF在体外的促有丝分裂作用有反应。然而,由于前列腺细胞分泌bFGF有限,bFGF介导的基质生长调节机制仍不清楚。