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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 promoter methylation of hsa-mir-34b (mir-34b) gene in early stage of non-small cell lung cancer (NSCLC). Methods The DNA of promoter region of mir-34b gene was amplified by nested PCR in 27 patients with stage Ⅰ NSCLC and the paired samples of paracancerous tissues. The variation of methylation profile was analyzed by cloning and sequencing And clinical and pathological features associated analysis. Results The methylation level of CpG island in cancer tissue mir-34b gene promoter was 9.7% (6.8% -22.3%), while it was 8.5% (5.3% -11.0%) in adjacent tissue. The methylation level of cancer tissue was higher than Paracancer tissue (Z = 2.355, P = 0.019). The incidence of methylation of the mir-34b promoter in smokers is higher than that of smokers and non-smokers. The level of methylation of mir-34b promoter in cancer tissue was not related to patient’s sex, age and PS score. Conclusion The methylation level of mir-34b promoter in NSCLC patients with early stage cancer is elevated. Smoking may be related to the methylation of mir-34b promoter. The mechanism of methylation of mir-34b associated with early oncosis deserves further study.