5-氮-2′-脱氧胞苷对人膀胱癌细胞系T_(24)细胞及其Apaf-1、APC基因甲基化状况的影响  

Effects of 5-Aza-CdR on proliferation and apoptosis of human bladder cancer T_(24) cells and expression of promoter methylation status of suppressor genes Apaf-1 and APC

作　　者：王文卫[1] 潘俊[2] 陈凌武[3] 林焕懿[1] 曾令友[1] 

机构地区：[1]中山大学附属第一医院黄埔院区泌尿外科,广东广州510700 [2]广东省中医院,广东广州510120 [3]中山大学附属第一医院泌尿外科,广东广州510080

出　　处：《重庆医学》2013年第13期1457-1459,共3页Chongqing medicine

基　　金：广东省科技计划基金资助项目(2009B060700044);广东省医学科研基金资助项目(A2005205);广州市黄埔区科技计划基金资助项目(330300409300403)

摘　　要：目的观察5-氮-2′-脱氧胞苷(5-Aza-CdR)对人膀胱癌T24细胞增殖、凋亡的影响及其对凋亡蛋白酶活化因子-1(Apaf-1)、结肠腺瘤性息肉病(APC)基因甲基化表达状况的影响。方法应用不同浓度的5-Aza-CdR处理人膀胱癌T24细胞,四甲基偶氮唑盐(MTT)比色法检测细胞生长抑制率;流式细胞术(FCM)检测细胞周期和凋亡的变化情况;以甲基化特异性PCR方法(MSP法)检测膀胱癌T24细胞株中Apaf-1及APC基因的甲基化状态。结果膀胱癌T24细胞中Apaf-1、APC基因分别呈半甲基化及高甲基化,5-Aza-CdR作用后T24细胞中Apaf-1及APC基因启动子区域均呈去甲基化状态;T24细胞的生长受到明显抑制,0.50、1.00、2.50、5.00μmol/L 5-Aza-CdR作用T24细胞72h后,细胞生长抑制率分别为0.50%、5.40%、20.40%、23.00%,组间比较差异有统计学意义(P<0.05),呈剂量和时间依赖关系。72h后G0/G1期的比例增加,S期比例明显减少,细胞凋亡率明显增加,5.00μmol/L 5-Aza-CdR组凋亡率为(40.35±3.63)%,对照组为(8.17±1.52)%,组间比较差异有统计学意义(P<0.01)。结论 5-Aza-CdR能逆转膀胱癌T24细胞中的Apaf-1及APC基因甲基化,恢复该基因的表达,诱导膀胱癌T24细胞凋亡。Objective To explore the effects of 5-Aza-2′-deoxycytidine（5-Aza-CdR） on the growth of T24 human bladder cancer cells and the expression of the promoter methylation status of the suppressor genes Apaf-1 and APC.Methods The methylation status of the promoter of Apaf-1 and APC gene in T24 cells were detected by the methylation-specific PCR（MSP）.Cultured T24 cells were treated with 5-Aza-CdR.MTT was used to detect the proliferation of T24 cells.Cell cycles and apoptosis changes were examined by means of flow cytometry（FCM）.Results The Apaf-1 and APC gene were semi-methylated or hypermethylated in T24 cells.5-Aza-CdR significantly inhibited the proliferation of bladder cancer cell in a time-and dose-dependent manner.Cultured T24 cells were treated with 0.50,1.00,2.50,5.00 μmol/L 5-Aza-CdR respectively.The inhibition was 0.50%,5.40%,20.40% and 23.00% respectively,which showing statistically significant difference（P0.05）.The cells in G1 phase and the proportion of apoptotic cells were significantly increased at 72 h after treatment with 5-Aza-CdR,while the cells in S phase were reduced.The proportion of apoptotic cells treated with 5.00 μmol/L 5-Aza-CdR was（40.35±3.63）%,and the control group was（8.17±1.52）%,with statistically significant difference（P0.01）.Conclusion 5-Aza-CdR may effectively induce the apoptosis of T24 cell through inhibiting the methylation of Apaf-1 and APC gene.

关 键 词：膀胱肿瘤 DNA甲基化 癌基因 5-氮-2′-脱氧胞苷 

分 类 号：R737.14[医药卫生—肿瘤]