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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 find out the difference of gene expression level and the expression level of related signaling pathways in the peripheral and transitional zone of prostate in healthy volunteers. Trying to explore the mechanism of the two in different proportions of prostate disease causes. Methods: Total RNA was extracted from the peripheral zone (PZ) and transition zone (TZ) fibroblasts by one step Trizol. The gene expression levels of CXCL1, CXCL2, CXCL3, IGFBP2, IGFBP3, IL-8, TAGLN, MEST were determined by quantitative RT-PCR. In addition, secreted proteins were collected and concentrated from PZ, TZ fibroblast conditioned media and the secreted proteins were compared by Western blotting. Results: The gene expression levels of some secretory proteins were significantly different between PZ and TZ fibroblasts. The mRNA expression of CXCL1, CXCL2, CXCL3 and IL-8 in TZ fibroblasts was significantly higher than that in PZ fibroblasts In the expression level. The mRNA expression of IGFBP2, IGFBP3 and TAGLN in PZ fibroblasts was significantly higher than that in TZ fibroblasts. The level of IGFBP3 and TAGLN protein secretion in PZ fibroblasts was significantly higher than that in TZ fibroblasts. Conclusion: The difference of gene expression and protein secretion between PZ and TZ fibroblasts may be involved in the significant difference between prostate cancer and benign prostatic hyperplasia (BPH) in different zones of prostate.