摘要
目的:探讨N-myc下游调节基因(N-myc downstream regulated gene-1,NDRG-1)基因在乳腺癌组织中的甲基化状态,研究甲基化酶抑制剂5-氮杂-2'-脱氧胞苷(5-Aza-2'-deoxycytidine,5-Aza-CdR)对乳腺癌细胞株T47D的生长增殖及NDRG-1mRNA表达的影响。方法:采用甲基化特异性PCR(methylation specific PCR,MSP)法检测乳腺癌组织、相应癌旁组织和乳腺良性病变组织中NDRG-1基因启动子甲基化状态;5-Aza-CdR处理T47D细胞后,MTT法观察细胞生长活性的变化,RT-PCR法检测处理前后抑癌基因NDRG-1的mRNA表达变化。结果:NDRG-1基因在乳腺癌组织中甲基化率为46.8%,癌旁组织中甲基化率为21.3%,而乳腺良性病变组织均未检测到甲基化。T47D细胞经5-Aza-CdR处理后,与对照组相比,细胞生长受到明显抑制。RT-PCR检测发现,与对照组相比,不同浓度处理组细胞的NDRG-1mRNA表达增多。结论:NDRG-1基因甲基化状态与乳腺癌发生有密切关系。5-Aza-CdR逆转T47D细胞NDRG-1基因甲基化,恢复该基因的表达,从而抑制肿瘤细胞生长。
Objective：To investigate the methylation status of N-myc downstream regulated gene-1（NDRG-1） gene in breast cancer and the effects of methylation enzyme inhibitor 5-Aza-2＇-deoxycytidine（5-Aza-CdR） on growth and expression of NDRG-1 mRNA in human breast cancer cell line T47D.Methods：Sensitive methylation-specific（MSP）-PCR was used to detect the methylation status in the promoter regions of NDRG-1 gene in 47 samples of breast cancer and tumor adjacent tissues and 15 cases of benign breast disease.The change in expression of the tumor suppressor gene NDRG-1 mRNA in cultured T47D cells was detected by RT-PCR before and after 5-Aza-CdR treatment.Cell proliferation was observed by MTT assay.Results：Hypermethylation frequencies of NDRG-1 gene promoter were 46.8% in breast cancer tissues and 21.3% in tumor adjacent tissues.No hypermethylation of NDRG-1 gene was observed in the tissues of breast benign disease.The growth of T47D cells was suppressed obviously after 5-Aza-CdR treatment compared with the control group.RT-PCR showed that compared with the control group,NDRG-1 mRNA expression was increased at different concentrations in 5-Aza-CdR treatment group.Conclusion：The promoter methylation status of NDRG-1 gene was significantly related with the occurrence of breast carcinoma.5-Aza-CdR could effectively reverse the methylation of NDRG-1 gene and recover its expression,thereby inhibiting the growth of tumor cells.
出处
《肿瘤》
CAS
CSCD
北大核心
2010年第4期310-313,共4页
Tumor
基金
青岛市公共领域科技支撑计划项目(编号:09-1-1-8-nsh)
