非小细胞肺癌TCF21基因表达及其启动子区甲基化的研究

摘要

目的 探讨非小细胞肺癌(NSCLC)中TCF21基因mRNA和蛋白的表达水平及其启动子区甲基化状态的意义.方法 运用RT-PCR、蛋白印迹法(Western-blot)和焦磷酸测序(Pyrosequencing)技术检测97例非小细胞肺癌组织和21例支气管扩张或肺大疱等手术切除的良性肺组织中TCF21基因mRNA、蛋白表达及启动子区甲基化状态.结果 97例NSCLC癌组织和21例良性肺组织中分别检测出TCF21基因mRNA阳性表达23例(23.71％)和14例(66.67％);NSCLC癌组织中TCF21蛋白平均表达量灰度值为0.49±1.78,良性肺组织中其平均表达量灰度值为1.48±1.58; NSCLC癌组织和良性肺组织中TCF21基因启动子区均有不同程度的甲基化,且癌组织和良性肺组织中各个位点甲基化频率均有统计学意义,在NSCLC癌组织中第1、5位点平均甲基化频率较高为49.04％和51.37％.结论 TCF21基因mRNA及蛋白表达在NSCLC癌组织和良性肺组织中均有统计学意义,TCF21基因启动子区平均甲基化频率明显高于良性肺组织细胞.%Objective Background and objective Bioinformatics technology found the TCF21 gene has difference expression in non-small cell lung cancer (NSCLC) and benign lung tissue.To explore TCF21 mRNA and protein expression in nonsmall cell lung cancer and its methylation of the promoter region is the aim of this study.Methods Use RT-PCR,Western blot and Pyrosequencing detected TCF21 gene mRNA,protein and the methylation of the promoter region respectively in 97 cases of non-small cell lung cancer and 21 cases of benign lung tissue.Results TCF21 gene mRNA-positive expression were detected 23 cases (23.71％) and 14 cases (66.67％) in 97 cases of NSCLC cancer tissue and 21 cases of benign lung tissue,the average gray value of TCF21 protein expression levels in NSCLC cancer tissue is 0.49 ± 1.78,while it is 1.48 ± 1.58 in benign lung tissue,the TCF21 gene promoter region have varying degrees methylation in NSCLC cancer tissue and benign lung tissue,and the significant of methylation frequency was found statistically significant between NSCLC cancer tissue and benign lung tissue,also it has higher frequency of 49.04％ and 51.37％ respectively at point 1 and 5 in NSCLC cancer organizations.Conclusion TCF21 gene mRNA and protein expression were statistically significant in NSCLC cancer tissue and benign lung tissue,the TCF21 gene promoter region average methylation frequency was significantly higher than that in benign lung tissue cells.