CAJ | 학술논문

提目的：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。
제목적：탐토5-잡담-2’-탈양포감（5-Aza-CdR）대인전렬선암세포（PC-3）주증식화조망적영향급기가능궤제。방법이2．5、5．0、10．0μmol/L적5-Aza-CdR작용우 PC -3세포48h후，채용류식세포술（FCM ）검측세포조망솔；역전록취합매련식반응（RT -PCR ）검측UCHL1 mRNA표체；갑기화측서취합매련반응（BSP）검측UCHL1 CpG도적갑기화상태；단백질인적법（Western Blot）검측UCHL1단백적표체。결과：5-Aza-CdR대PC-3세포생장유억제작용；여대조조상비세포조망솔명현증고（ P＜0．01）；5-Aza -CdR처리세포후UCHL1적계동자갑기화수평명현강저（ P＜0．01）；UCHL1 mRNA표체수평현저상조（ P＜0．01）；UCHL1단백표체수평상승（ P＜0．01）。결론：5-Aza -CdR능유도전렬선암PC-3세포주조망적작용，기궤제가능시5-Aza-CdR능역전PC -3세포UCHL1계동자CpG도적이상갑기화，유도mRNA전록화단백적표체。
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 (FCM);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 UCHL1 protein were determined using Western Blot technique .Results The apoptosis rate of 5-Aza-CdR treated with PC -3 cells was significantly higher than control (P<0 .01) . UCHL1 promoter methylation levels decreased dramatically after the cells were treated with 5 - 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 UCHL 1 CpG island ,promote the expression of UCHL 1 RNA and protein .