5-杂氮-2’-脱氧胞苷调控UCHL1基因表达对人前列腺癌PC-3细胞增殖及凋亡的影响

目的：探讨5-杂氮-2’-脱氧胞苷（5-Aza—CdR）对人前列腺癌细胞（PC-3）株增殖和凋亡的影响及其可能机制。方法以2．5、5．0、10．0kenol／L的5-Aza—CAR作用于PC-3细胞48h后，采用流式细胞术（FCM）检测细胞凋亡率；逆转录聚合酶链式反应（tiT—PcR）检测UCHL1 mRNA表达；甲基化测序聚合酶链反应（BsP）检测UCHLlCpG岛的甲基化状态；蛋白质印迹法（Western Blot）检测UCHL1蛋白的表达。结果：5-Aza—CAR对PC-3细胞生长有抑制作用；与对照组相比细胞凋亡率明显增高（P〈0．01）；5-Aza—CdR处理细胞后UCHL1的启动子甲基化水平明显降低（P〈0．01）；UCHL1 mRNA表达水平显著上调（P〈0．01）；UCHL1蛋白表达水平上升（P〈0．01）。结论：5-Aza—CAR能诱导前列腺癌PC-3细胞株凋亡的作用，其机制可能是5-Aza—CdR能逆转PC-3细胞UCHLl启动子CpG岛的异常甲基化，诱导mRNA转录和蛋白的表达。

Effects of 5 - Aza - 2＇ - deoxycytidine on proliferation and apoptosis by modulating U CHL1 gene expression in human prostate cancer cell line PC- 3

Objective：To investigate the effects of 5- Aza- 2＇ -deoxycytidine（5 - Aza- CdR） on the proliferation and apoptosis in human prostate cancer （PCa） PC - 3 cells and the possible mechanism. Methods：The PC - 3 cells were treated with 5 - Aza- CdR at concentrations of 2. 5, 5.0, 10.0 uLmol/L for 48h. Their apoptosis rate was detected by Flow cytometry （ FCMI ; CpG island of ubiquitin carboxyl - terminal hydrolase L1 { UCHL1 } methylation levels were determined by Bisulfite sequencing PCR{ BSP）. UCHL1 mRNA expression levels were determined via RT- PCR. whereas the expression of UCI-IL1 protein were determined using Western Blot technique. Results The apoptosis rate of 5 - Aza - CdR treated with PC - 3 ceils was significantly higher than control（ P 〈 0.01 ）. UCHL1 promoter methylation levels decreased dramatically after the cells were treated with5 - Aza - CdR （ P 〈 0.01 ）. The mRNA expression of UCHL1 dramatically increased（ P 〈 0.01 }. The protein expression of UCHL1 significantly increased（ P 〈 0.01 ）. Conclusions 5 - Aza - CdR can induce the apoptosis of PC - 3 prostate cancer cell line, the mechanism may be 5 - Aza - CdR can reverse the methylation of UCI-IL1 CpG island, promote the expression of UCHL1 RNA and protein.