前列腺增生间质对上皮细胞组织激肽释放酶7表达的影响

Stromal cells inhibit Kallikrein 7 expression in the epithelial cells of benign prostate hyperplasia

目的:研究前列腺增生间质细胞对上皮细胞组织激肽释放酶7(KLK7)表达的影响。方法:分离良性前列腺增生(BPH)组织的间质细胞与上皮细胞,经细胞形态学及化学发光微粒子免疫分析(CM IA)方法证实后,构建上皮/间质细胞共培养模型;分别提取单独、共培养条件下上皮细胞的mRNA与蛋白;RT-PCR检测KLK7mRNA表达变化,W estern印迹检测KLK7基因编码蛋白(hK7)表达的改变。结果:细胞形态学、CM IA结果显示成功分离上皮与间质细胞,并构建前列腺间质/上皮细胞共培养模型;RT-PCR检测发现,KLK7基因mRNA的表达在有间质细胞共培养的上皮细胞中均高于单独培养的上皮细胞。KLK7/GAPDH的灰度比值分别为1.41±0.04、1.78±0.10,具有显著差异(P<0.01);W estern印迹与RT-PCR结果一致。结论:前列腺增生间质细胞抑制上皮细胞KLK7基因表达,KLK7可能是前列腺上皮细胞应对间质细胞调控的一种效应物质。

Objective： To investigate the influence of stromal cells on the Kallikrein 7 （KLK7） expression of epithelial cells in benign prostate hyperplasia （BPH）. Methods ： We constructed a stromal-epithelial co-culture model after separating the two types of cells from BPH tissues and identifying them by cell morphology and chemiluminescent microparticle immunoassay （CMIA）. The expression of KLK7 mRNA was detected by RT-PCR in the epithelial cells with or without the stromal cells, and that of the KLK7 protein （hK7） determined by Western blot. Results： Stromal and epithelial cells were successfully separated and identified, and a stromal-epithelial co-culture model successfully established. RT-PCR showed that the mRNA expression of the KLK7 gene was higher in the epithelial cells co-cultured with stromal cells than in the epithelial cells alone, and the gray value of KLK7 to GAPDH was 1.41 ±0.041 in the former and 1.78 ±0.10 in the latter （P 〈 0.01 ）. The results of Western blot were consistent with those of RT-PCR. Conclusion： Stromal cells can suppress the expression of the KLK7 gene in the epithelial cells in BPH. KLK7 may be involved in the change of epithelial cells stimulated by stromal cells.