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目的 研究良性前列腺增生上皮细胞在有无间质细胞共培养条件下的基因差异表达。 方法 利用前列腺间质与上皮细胞共培养模型及DDRT PCR技术 ,对单独培养的前列腺增生上皮细胞和与间质细胞共培养的前列腺增生上皮细胞的mRNA进行差异表达分析 ,获得差异表达片段(ESTs) ;并对这些ESTs进行反向Northern印迹分析证实及克隆测序 ,将所测阳性克隆的cDNA序列与核苷酸数据库中已知序列进行同源性比较。 结果 得到ESTs 70个 ;经同源性分析 ,4 5个ESTs与已知基因有较高同源性 ,2 5个ESTs为新的cDNA片段。 结论 良性前列腺增生上皮细胞在有无间质细胞共培养条件下 ,存在差异表达基因 ,这些差异基因可能与前列腺间质细胞对前列腺增生上皮细胞的作用有关。
Objective To study the differential gene expression of benign prostatic hyperplasic epithelial cells in the presence or absence of mesenchymal cells co-culture. Methods Differential expression of mRNA of prostatic hyperplasia epithelial cells cultured in vitro and prostatic hyperplasia epithelial cells co-cultured with mesenchymal cells was studied by using the model of prostatic mesenchymal and epithelial cells co-culture and DDRT PCR technique to obtain differentially expressed fragments (ESTs) ; Reverse Northern blot analysis of these ESTs was carried out to confirm and clone sequencing. The cDNA sequences of the positive clones were compared with the known sequences in the nucleotide database. The results obtained 70 ESTs; by homology analysis, 45 ESTs had high homology with known genes, 25 ESTs were new cDNA fragments. Conclusion There are differentially expressed genes in benign prostatic hyperplasic epithelial cells with and without mesenchymal cells co-culture. These differential genes may be related to the role of prostatic stromal cells in prostatic hyperplasia epithelial cells.