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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。
Objective: To investigate the mechanism of DLC-1 gene low expression in MCF-7 human breast cancer cell line. Methods: Methylation-specific PCR (MSP) was used to detect the methylation status of DLC-1 gene in human breast cancer cell line MCF-7. Different concentrations of 5-Aza-2’-deoxycytidine ) Was used to detect the expression of DLC-1 mRNA in human breast cancer cell line MCF-7 before and after treatment with RT-PCR and Real-time PCR. Results: The CpG island in the promoter region of DLC-1 gene was methylated. After 5-Aza-CdR treatment, the promoter region of DLC-1 gene was demethylated and its mRNA was restored. CONCLUSION: Methylation of the tumor suppressor gene DLC-1 CpG island is one of the reasons leading to the low expression of this gene. 5-Aza-CdR can reverse the methylation status of DLC-1 gene.