摘要
目的 检测早期非小细胞肺癌（NSCLC）患者癌组织与癌旁组织hsa-mir-34b（mir-34b）基因启动子甲基化水平的差异.方法 采用重亚硫酸盐还原技术、降落巢式PCR扩增27例Ⅰ期NSCLC患者癌组织和癌旁组织配对样本中mir-34b基因启动子区DNA,克隆测序法分析甲基化谱变异,并与临床病理特征进行关联分析.结果 癌组织mir-34b基因启动子CpG岛整体甲基化水平为9.7%（6.8%～22.3%）,而癌旁组织为8.5%（5.3%～11.0%）,癌组织甲基化水平高于癌旁组织（Z=2.355, P=0.019）.当前吸烟患者mir-34b基因启动子甲基化发生率高于以往吸烟及不吸烟者.癌组织mir-34b基因启动子甲基化水平与患者性别、年龄、PS评分无关.结论 早期NSCLC患者癌组织mir-34b基因启动子甲基化水平升高,吸烟可能与mir-34b基因启动子甲基化的发生有关.mir-34b基因甲基化与早期肿瘤发生相关的机制值得进一步研究.
Objective To detect the methylation status of mir-34b gene promoter, and analyze the different methylation levels in tumor tissues and adjacent tissues in early stage non-small cell lung cancer （NSCLC） patients. Methods After bisulfite treatment of genome DNA, touchdown-nest PCR was performed to amplify the promoter CpG islands of mir-34b gene in paired tumor and adjacent tissues from 27 surgically resected stage I NSCLC patients, and the methylation status of mir-34b gene promoter was detected by clo- ning-based sequencing method. Then the correlation between methylation level and clinical factors of patients was analyzed. Results The methylation level of mir-34b gene promoter in tumor tissues was 9. 7% （6. 8-22. 3% ）, while it＇s 8. 5% （5.3-11. 0% ）in adjacent tissues. The differences between tumor and adjacent tissues were statistically significant （ Z = 2. 355, P = 0. 019）. The methylation level in tumor tissues was correlated with smoking status. Conclusion Methylation level of mir-34b gene in NSCLC tissues was higher than its in adjacent tissues, which was also correlated with smoking status, suggesting a potential relationship between the tumorigenesis in early stage and the methylaiton of mir-34b gene promoter.
出处
《临床肿瘤学杂志》
CAS
2010年第11期961-965,共5页
Chinese Clinical Oncology
基金
2010年度广东省医学科学研究基金(A2010011)
