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甲基双加氧酶对非小细胞肺癌增殖、迁移和侵袭的影响及机制

Effects of ten-eleven translocation-2 on proliferation and migration as well as invasion of non-small cell lung cancer and its mechanism

    摘要
  • 摘要:
      目的  探讨甲基双加氧酶(TET2)对非小细胞肺癌(NSCLC)细胞增殖、迁移和侵袭能力的影响及其作用机制。
      方法  通过在线数据库分析TET2与NSCLC患者预后的相关性；实时荧光定量聚合酶链反应(qRT-PCR)、Western blot检测人正常支气管上皮细胞及非小细胞肺癌系中TET2的表达；瞬时转染si-TET2干扰序列，运用CCK8、克隆形成法检测细胞的增殖；Transwell法检测细胞的迁移和侵袭；流式细胞术检测细胞周期变化；Western blot检测上皮间质转化(EMT)标志物以及N-cadherin、E-cadherin、Vimentin、Cyclin D1蛋白水平。
      结果  TET2高表达的患者总体生存率差，相比于人正常肺上皮细胞，TET2在NSCLC细胞株中高表达。敲低TET2可显著抑制A549、H1299细胞的增殖、迁移和侵袭(P < 0.001)，并将细胞周期阻滞于G0/G1期。Western blot结果显示，N-cadherin、Vimentin和Cyclin D1蛋白表达显著下调，E-cadherin表达显著上调(P < 0.05或P < 0.01或P < 0.001)。
      结论  TET2可能通过影响细胞周期以及EMT的关键蛋白促进NSCLC的增殖、迁移和侵袭。

     

    Abstract:
      Objective  To discuss the effect of ten-eleven tanslocation-2 (TET2) on the proliferation, migration and invasion of non-small cell lung cancer (NSCLC) and its mechanism.
      Methods  Online databases were used to detect the correlation between TET2 expression and prognosis of NSCLC patients; quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were performed to detect the expression of TET2 bronchial epithelial cells and non-small cell lung cancer lines; after transient transfection with si-TET2, cell proliferation were detected by CCK8 and clonogenic assay; Transwell assay was used to detect cell migration and invasion; cell cycle changes were detected by flow cytometry; epithelial-mesenchymal transition (EMT) markers and protein levels of N-cadherin, E-cadherin Vimentin and Cyclin D1 were detected by Western blot.
      Results  Patients with high TET2 expression had poor overall survival. Compared with normal human lung epithelial cells, TET2 is highly expressed in NSCLC cell lines. TET2 knockdown significantly inhibited the proliferation, migration and invasion of A549 and H1299 cells (P < 0.001), and blocked the cell cycle in the G0/G1 phase. Western blot results showed that the protein expressions of N-cadherin, Vimentin and Cyclin D1 were significantly down-regulated, while the expression of E-cadherin was significantly up-regulated (P < 0.05, P < 0.01 or P < 0.001).
      Conclusion  TET2 may promote the proliferation, migration and invasion of NSCLC by affecting the cell cycle and key proteins of EMT.

     

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