5氮杂脱氧胞苷对乳腺癌MCF-7细胞中抑癌基因p16 mRNA表达的影响  

Effect of 5-aza-2 '-deoxycytidine on demethylation of p16 gene in breast cancer cell line MCF-7

作　　者：叶静[1] 范昭 胡慧 李明华[1] 万汇涓[1] 于洁[1] YE Jing FAN Zhao HU Hui LI Ming hua WAN Hui juan YU Jie 

机构地区：[1]北京大学深圳医院 中心实验室 [2]腺体外科,广东深圳518036 [3]Central Laboratory [4]Department of Gland Surgery ,Peking University Shenzhen Hospital ,Shenzhen 518036,China

出　　处：《临床荟萃》2012年第6期489-491,494,共4页Clinical Focus

基　　金：深圳市科技计划项目(201103029)

摘　　要：目的探讨5氮杂脱氧胞苷(5-aza-cdr)对乳腺癌细胞中p16基因mRNA表达的影响。方法分别用5-aza-cdr 5、10和20μmol/L处理乳腺癌细胞MCF-7,未处理的MCF-7细胞作为对照。采用甲基化特异性聚合酶链反应(MSP)对药物处理前后的细胞进行p16基因甲基化检测;绿色荧光染料实时反转录聚合酶链反应(SYBR GreenqRT-PCR)检测p16mRNA表达。结果在未处理的MCF-7细胞中(对照)p16基因呈完全甲基化状态,随着5-aza-cdr浓度增加,甲基化水平逐渐减弱,至5-aza-cdr 20μmol/L时p16基因甲基化状态完全被逆转;p16mRNA表达水平也随着药物剂量的递增逐渐增加,5-aza-cdr 20μmol/L处理组与5、10μmol/L处理组以及未处理组比较差异均有统计学意义(均P<0.01)。结论乳腺癌细胞MCF-7中p16基因的甲基化能被5-aza-cdr逆转,去甲基化后可以促进p16mRNA表达。Objective To investigate the effect of 5-aza-2 ＇-deoxycytidine（5-aza-cdr） on p16 gene demethylation mRNA expression in breast cancer cell. Methods MCF-7 cells were treated with 5-aza-cdr 5,10,20 umol/L and the untreated MCF-7 cell served as control. Methylation and mRNA levels for p16 gene were analyzed by methylation- specific polymerase chain reaction（MSP） and SYBR Green qRT-PCR. Results p16 gene was completely methylated in the control group. After culturing with the increased 5-aza-cdr, methylation levels in MCF-7 cells were gradually weakened and the completely reversed methylation occurred in the concentration 20 umol/L, p16 mRNA levels were stepped up with 5-aza-cdr increasing. This mRNA level in 20 /~mol/L treated group showed significant improvement in comparison with 5,10 umol/L treated group as well as control group （ P d0.01）. Conclusion The methylation of p16 gene can be reversed in MCF-7 cell treated by 5-aza-cdr. This demethylation can promote expression of p16 gene mRNA.

关 键 词：乳腺肿瘤 DNA甲基化 基因 p16 聚合酶链反应 

分 类 号：R737.9[医药卫生—肿瘤]