| 摘要: |
| 目的 研究三叶因子1（TFF1）在肺癌组织及肺癌细胞株中的表达、甲基化状态及其功能。方法 选取98 例非小细胞肺癌患者手术切除的肺癌组织，采用RT-PCR、MSP 检测TFF1 在肺癌组织中的表达与甲基化状态，分析其与患者临床特点的相关性，以及检测肺癌细胞株H23、H1299、L78、H446、H157 及95D 中TFF1 的表达和甲基化状态以及5-aza-2'-deoxycytidine（DAC）处理后细胞株中TFF1 表达的变化。TFF1 转染H23 细胞株，MTT 法及克隆形成实验检测其增殖能力和克隆能力的变化，细胞凋亡实验及Western blot 检测凋亡相关蛋白Caspase3 表达水平。结果 TFF1 在肺癌组织中的甲基化率为56.1%（55/98）。TFF1 甲基化与肺癌患者的TNM 分期和淋巴结转移显著相关（P＜0.001）。TFF1 在H23 和H157 中无表达，在H1299、H446 及95D 中表达较低，在L78 中表达较高。DAC 处理后，细胞株TFF1 表达的变化分别为表达恢复（H23、H157），表达增强（H1299，H446，95D）和无明显改变（L78）。恢复表达TFF1 后，H23 细胞株的增殖及克隆形成能力明显减弱（P＜0.05），细胞凋亡率和凋亡相关蛋白Caspase3 活性片段的表达明显增加。结论 TFF1 在肺癌组织中的甲基化状态与肺癌的TNM 分期和转移密切相关。TFF1 在肺癌细胞中的表达受DNA 甲基化的调控。肺癌细胞株H23 中恢复表达TFF1 可以抑制其增殖和克隆能力，促进其凋亡。 |
| 关键词: TFF1 肺癌 DNA甲基化 表观遗传学 |
| DOI： |
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| Expression and methylation status of TFF1 in lung cancer |
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LIU Kaitai, LU Miaozhen, CHU Yudong
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Ningbo Medical Center
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| Abstract: |
| Objective To investigate the expression and methylation status of Trefoil factor 1(TFF1) in lung cancer tissues and lung cancer cell lines. Methods Expression and methylation status of TFF1 in 98 non-small cell lung cancer (NSCLC) tissues and lung cancer cell lines (H23, H1299, L78, H446, H157 and 95D) were detected by RT-PCR and methylation specific PCR (MSP) , respectively. The changes of TFF1 expression was also examined after the lung cancer cell lines treated with 5-aza-2'-deoxycytidine(DAC). After H23 cells were transfected with TFF1 gene, the variation of proliferation and cloning ability of H23 cells was examined with MTT assay and colony formation, respectively. The apoptosis H23 cells was also determined and the expression of Caspase3 was detected with Western blot after transfection. Results The methylation rate of TFF1 in lung cancer tissues was 56%(55/98). TFF1 methylation was significantly correlated with TNM stage and lymph node metastasis in lung cancer patients (P＜0.001). There was no expression of TFF1 in H23 and H157 cells, low expression in H1299, H446 and 95D cells, and high expression in L78 cells; after treatment of DAC, the expression of TFF1 was restored (H23 and H157), increased (H1299, H446 and 95D) or not changed (L78). Transfection of TFF1 decreased proliferation and colony formation, promoted cell apoptosis rate and enhanced expression of Caspase3 in H23 cells. Conclusion The methylation status of TFF1 in NSCLC tissue is closely related to TNM stages and metastasis of lung cancer. The expression of TFF1 in lung cancer is associated with DNA methylation. Restoration of TFF1 expression in lung cancer cells may suppress cell proliferation and accelerate cell apoptosis. |
| Key words: TFF1 Lung cancer DNA methylation Epigenetic |
