| 宗 丹,郭文杰,王德军.ZNF488基因对鼻咽癌细胞HONE1生物学行为的影响[J].肿瘤学杂志,2014,20(2):115-121. |
| ZNF488基因对鼻咽癌细胞HONE1生物学行为的影响 |
| Effects of ZNF488 on the Biological Behavior of Nasopharyngeal Carcinoma HONE1 Cell Line |
| 投稿时间：2013-11-11 |
| DOI：10.11735/j.issn.1671-170X.2014.02.B008 |
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| 中文关键词: 鼻咽肿瘤 锌指蛋白488 上皮—间质转化 侵袭转移 分子机制 |
| 英文关键词:nasopharyngeal neoplasms ZNF488 EMT invasion and metastasis molecular mechanism |
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| 中文摘要: |
| 摘 要：［目的］ 探讨ZNF488基因对鼻咽癌细胞HONE1生物学行为的影响。［方法］ 构建携带空载体、人ZNF488基因的逆转录病毒质粒pMSCV和pMSCV-ZNF488，以磷酸钙法转染293FT细胞并收集病毒液，感染鼻咽癌细胞株HONE1，经嘌呤霉素筛选及Western Blot鉴定，建立稳定表达ZNF488的HONE1-pMSCV-ZNF488和空载体HONE1-pMSCV的细胞株。应用噻唑盐（MTT）法、平板克隆形成实验检测ZNF488过表达对HONE1细胞生长增殖的影响，同时应用流式细胞术检测ZNF488对细胞周期增殖指数的影响。划痕实验及Transwell体外侵袭实验观察ZNF488对HONE1迁移侵袭能力的影响。Western Blot检测上皮—间质转化（EMT）相关蛋白及抑癌基因PTEN的表达情况。［结果］ 成功建立稳定表达ZNF488的HONE1-pMSCV-ZNF488细胞株，MTT 法及平板克隆形成实验结果显示，与空载体组HONE1-pMSCV细胞相比，HONE1-pMSCV-ZNF488细胞增殖及克隆形成能力均增强。流式细胞术显示过表达ZNF488细胞 S期百分比较空载体组高，增殖指数增加。划痕实验、Transwell 体外侵袭实验示ZNF488能够显著增加细胞的迁移、侵袭能力。Western Blot检测发现上皮标志E-Cadherin、α-Catenin表达降低，间质性标志Fibronectin、Vimentin表达升高，同时抑癌因子PTEN表达下降。［结论］ 过表达ZNF488可通过诱导鼻咽癌细胞HONE1发生上皮间质转变及下调抑癌基因PTEN，促进细胞的增殖及迁移侵袭能力。 |
| 英文摘要: |
| Abstract：［Purpose］ To investigate the effects of zinc finger protein 488(ZNF488) on the biological behavior of nasopharygeal carcinoma (NPC) HONE1 cell line. ［Methods］ Retrovirus vector pMSCV or pMSCV-ZNF488 was transfected into 293FT cells to produce retrovirus，then the nasopharyngeal carcinoma cell line HONE1 was infected to construct ZNF488-expressing stable cell line and vector control group. The cells were screened by puromycin. MTT assay，colony formation assay and flow cytometry cell cycle analysis were applied to evaluate the proliferation in both HONE1-ZNF488 cells and the control vector. Wound and healing assay and Transwell assay were performed to analyze the effect on cell migration and invasion. Western Blot was used to detect protein levels of EMT markers and suppressor gene PTEN.［Results］ After stable cell lines expressing ZNF488 and the control vector were successfully established，MTT assay and colony formation assay showed that overexpression of ZNF488 significantly enhanced cell proliferation compared with control group. The flow cytometer analysis result indicated ZNF488 might accumulated cells in S phase. Wound and healing and Transwell assay showed that ZNF488-expressing cells endowed the cells with much more migratory and invasive properties than the vector. Mechanistically，we found that overexpression of ZNF488 lead to the down-regulation of epithelial markers E-Cadherin and α-Catenin and up-regulation of mesenchymal markers Fibronectin and Vimentin，as well as the downregulation of tumor suppressor gene PTEN. ［Conclusion］ ZNF488 might promote potentiation of growth，proliferation，invasion and metastasis of HONE1 cell line，which might be associated with induced EMT and downregulation of suppressor gene PTEN. |
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