DLC-1 基因在MCF-7 人乳腺癌细胞系中低表达的机制探究

目的:探究DLC-1基因在MCF-7人乳腺癌细胞系中低表达的机制。方法:应用甲基化特异性PCR(MSP)检测人乳腺癌细胞MCF-7的DLC-1基因甲基化状态,不同浓度的5-氮杂-2’-脱氧胞嘧啶(5-Aza-CdR)处理人乳腺癌细胞MCF-7,RT-PCR及Real-time PCR定量检测用药前后细胞中DLC-1基因mRNA表达水平变化。结果:DLC-1基因启动子区CpG岛呈甲基化状态,经过5-Aza-CdR处理后,DLC-1基因启动子区呈去甲基化状态,并且其mRNA恢复表达。结论:抑癌基因DLC-1 CpG岛甲基化是导致该基因低表达的原因之一,5-Aza-CdR能逆转DLC-1基因甲基化状态。

Study on Mechanism of Low Expression of Tumor Suppressor GeneDLC-1 in Human Breast Cancer Cell Line

Objective: To investigate the mechanism of low expression of tumor suppressor gene dlc-1 in human breast cancer cellline. Methods:Methylation-specific PCP (MSP) was used to examine the methylation status of DLC-1 promoter.MCF-7 cells were treatedby different concentrations of 5-Aza-CdR. DLC-1 mRNA levels were examined by RT-PCR and Real-time PCR before and after treatmentwith 5-Aza-CdR. Results: There was methylation in DLC-1 promoter. The expression of DLC-1 mRNA in MCF-7 increased after treatedby by 5-Aza-CdR. Conclusion:Methylation of DLC-1 promoter might be the main reason of the low expression of DLC-1 inMCF-7 humanbreast cancer cells, and the 5-Aza-CdR can inhibit this methylation.