正常人前列腺外周带和移行带成纤维细胞差异性基因表达与蛋白分泌的研究

Study on the Differences of Gene Expression and Protein Secretion Level of the Normal Prostate Peripheral Zone and Transitional Zone Fibroblast Cells

目的:寻找正常人前列腺外周带和移行带细胞在基因表达水平及相关信号通路蛋白表达水平的差异。试图从机理上探求两者在不同前列腺疾病中所占比例不同的原因。方法:用Trizol一步法从外周带(PZ)和移行带(TZ)成纤维细胞中抽提总RNA。通过定量RT-PCR技术测定CXCL1、CXCL2、CXCL3、IGFBP2、IGFBP3、IL-8、TAGLN、MEST的基因表达水平。此外,从PZ、TZ成纤维细胞条件培养基中收集、浓缩分泌的蛋白,通过Western blotting技术比较分析分泌蛋白的水平。结果:一些分泌蛋白的基因表达水平在PZ与TZ成纤维细胞存在显著差别,其中CXCL1、CXCL2、CXCL3与IL-8的mRNA在TZ成纤维细胞中的表达水平明显高于其在PZ成纤维细胞中的表达水平。而IGFBP2、IGFBP3与TAGLN的mRNA在PZ成纤维细胞中的表达水平明显高于其在TZ成纤维细胞中的表达水平。IGFBP3与TAGLN在PZ成纤维细胞中的蛋白分泌水平明显高于其在TZ成纤维细胞中的蛋白分泌水平。结论:PZ与TZ成纤维细胞中基因表达及蛋白分泌水平的差异可能参与导致前列腺癌与前列腺增生(BPH)发病率在前列腺不同区带的显著区别。

Objective：To detect the different expression levels of gene and protein in revelent signal pathway between the peripheral zone（PZ） and transitional zone（TZ） of human normal prostate and to find the reason of different proportions in the different prostate diseases about the mechanism between the two cell lines.Methods：Total RNA with Trizol was extracted from the PZ and TZ fibroblast cells.The expression level of gene was determinated by Real-time PCR technology.Secretory proteins were collected and concentrated from the conditioned medium of the two fibroblast cells.The level of the secretory protein was contrasted and analyzed by Western blotting.Results：There were significant differences about the expression level of some genes between the PZ and TZ fibroblast cells.Levels of four gene expression including CXCL1,CXCL2,CXCL3 and IL-8 in TZ fibroblasts were significantly higher than in PZ fibroblast cells.While the expression levels of three genes IGFBP2,IGFBP3 and TAGLN in PZ fibroblasts were significantly higher than those in TZ fibroblast cells.Finally,the protein secretion levels of IGFBP3 and TAGLN in PZ fibroblast cells is significantly higher than in TZ fibroblast cells.Conclusion：There are some differences in gene expression and protein secretion between the PZ and TZ fibroblast cells.It may be involved in the cause of the remarkable distinction about the morbidity of the prostate cancer and benign prostatic hyperplasia（BPH） in different zones of of prostate.