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  

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

作　　者：李俊枫[1] 罗子国[1] 丁璇[1] 唐溧 

机构地区：[1]重庆医科大学生命科学院,重庆400016

出　　处：《激光杂志》2013年第6期107-109,共3页Laser Journal

摘　　要：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、10.0μmol/L的5-Aza-CdR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(RT-PCR)检测UCHL1 mRNA表达;甲基化测序聚合酶链反应(BSP)检测UCHL1 CpG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-CdR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-CdR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UCHL1启动子CpG岛的异常甲基化,诱导mRNA转录和蛋白的表达。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.

关 键 词：5-杂氮-2’-脱氧胞苷 UCHL1 甲基化 

分 类 号：TN244.8[电子电信—物理电子学] R737.25[医药卫生—肿瘤]