MEST基因在正常人前列腺移行带和外周带基质成纤维细胞中的mRNA表达差异及其与DNA甲基化水平的关系

Methylation and mRNA Expression of MEST Gene in Normal Prostate Peripheral Zone and Transitional Zone Fibroblast Cells

目的:探讨正常人的前列腺移行带(PZ)和外周带(TZ)基质成纤维细胞中MEST(mesoderm specific transcript)基因的甲基化水平变化和表达差异,及DNA甲基化抑制剂(5-Aza-CdR)对其表达水平的影响。方法:用硫化测序PCR(bisulfite sequencing PCR,BSP)和实时荧光定量PCR方法分别检测5-Aza-CdR处理前、后在前列腺PZ和TZ原代成纤维细胞中MEST基因的甲基化水平和相应的mRNA表达。结果:MEST基因在PZ成纤维细胞中为低甲基化高表达,在TZ成纤维细胞中为高甲基化低表达。加入5-Aza-CdR后,MEST在PZ和TZ成纤维细胞中都为去甲基化,而表达水平上升,但是TZ成纤维细胞表达水平的改变比PZ成纤维细胞大。结论:MEST基因的DNA甲基化差异可能是前列腺外周带和移行带成纤维细胞生物学行为差异的分子基础。

Objective： To investigate the methylation and the expression level of mesoderm specific transcript （MEST） gene between the prostate peripheral zone （PZ） and transitional zone （TZ） fibroblast cells of human normal prostate, and to explore the effect ofDNA methylation inhibitor on its expression. Methods： Bisulfite sequencing PCR （BSP） and Real-time PCR were used to detect MEST methylation and mRNA expression respectively in PZ and TZ fibroblast cells before and after treatment with DNA methylation inhibitor （5-Aza- CdR）. Results： The methylation level of MESTwas lower and the mRNA level of MESTwas higher in PZ fibroblast cells, compared with those in TZ fibroblast cells. After being treated with 5-Aza-CdR, both cells showed hypomethylation and up-regulated expression of MEST gene. However, the MEST mRNA expression in TZ fibroblast cells was significantly higher than that in PZ fibroblast cells. Conclusion： DNA methylation of MEST gene may be the molecular basis of different biological behaviors of fibroblast cells in prostate PZ and TZ.