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目的研究正常前列腺移行带和外周带在基因表达上存在的差异。方法二步法提取移行带和外周带组织总RNA,应用含有6700条人cDNA微距阵基因芯片,分别检测17例正常前列腺移行带和外周带的基因表达谱并重复3次,用cy5dUTP标记移行带组织mRNA,cy3dUTP标记周围带组织mRNA。Ratio均数(RA)<0.5或>2入选为表达差异的基因,RA<0.2或>4判断为表达有极显著差异的基因。采用RT PCR法验证差异表达的基因。结果移行带和外周带有表达差异的基因共640条(移行带294条,外周带346条),在GeneBank中功能明确者371条。表达有极显著差异的基因14条,其中移行带高表达的有表皮生长因子(EGF),外周带高表达的有成骨细胞刺激因子1(OSF1)。RT PCR结果证明在17例正常前列腺移行带和20例良性前列腺增生尿道周围组织中,EGF与内参基因β肌动蛋白的相对表达量明显高于外周带。结论正常前列腺移行带和外周带存在着基因表达差异,这可能是两带性结构差异生物学行为的分子基础;EGF对移行带的生长有重要促进作用。
Objective To investigate the differences in gene expression between normal and peripheral bands of normal prostate. Methods Total RNA was extracted from the transition zone and peripheral zone by two-step method. Gene expression profiling of 17 normal prostate transitional zone and peripheral zone was detected by using 6700 human cDNA microarray microarray. The gene expression profile was repeated 3 times and transfered with cy5dUTP marker Tissue mRNA, cy3dUTP marker around the tissue mRNA. Ratio mean (RA) <0.5 or> 2 was selected as a gene for differential expression, and RA <0.2 or> 4 was judged as a gene with very significant difference. The differentially expressed genes were verified by RT PCR. Results A total of 640 genes (transitional zone and 346 peripheral zone) were found in transition zone and peripheral zone, and 371 genes were identified in GeneBank. There were 14 genes with extremely significant differences, including epidermal growth factor (EGF) with high expression in transitional zone and osteoclast stimulating factor 1 (OSF1) with highly expressed in peripheral zone. The results of RT-PCR showed that the relative expression of EGF and β-actin was significantly higher than that in the peripheral zone in 17 cases of normal prostate and 20 cases of benign prostatic hyperplasia. Conclusion There is a difference in gene expression between normal and peripheral bands in normal prostate, which may be the molecular basis of the difference in biological behavior between two bands. EGF may play an important role in promoting the growth of transitional bands.