hsa—miR-218下调Survivin表达影响鼻咽癌CNE-2Z细胞增殖和凋亡

hsa-miR-218 Down-Regulates Survivin and Contributes to Proliferation and Apoptosis in Nasopharyngealeareinoma CNE-2Z Cells

目的研究hsa-miR-218对人鼻咽癌CNE-2Z细胞增殖和凋亡的影响，探讨其作用机制。方法构建hsa-miR-218的真核表达载体（pmiR-218），利用LipofectamineTM2000将PmiR-218转染CNE-2Z细胞，四甲基偶氮唑蓝（MTT）法检测PmiR-218对CNE-2Z细胞增殖的影响；AnnexinV／PI双染流式细胞仪检测，miR-218对CNE-2Z细胞凋亡的影响；Western印迹检测细胞内Survivin蛋白表达水平。结果成功构建hsa-miR-218的真核表达载体，miR-218。与随机对照组相比，转染PmiR-218的CNE-2Z细胞增殖受到明显抑制（P〈0．05），细胞早期凋亡显著增加（P〈0．05）。与随机对照组相比，PmiR-218抑制细胞内Survivin蛋白的表达。结论，miR-218可抑制人鼻咽癌CNE-2Z细胞增殖，诱导细胞凋亡，其作用机制与下调Survivin基因表达有关，提示hsa-miR-218在鼻咽癌的发生发展中扮演着肿瘤抑制基因的作用，可能是鼻咽癌治疗的新靶点。

Objective To investigate the effect of hsa-miR-218 on cell proliferation and apoptosis in nasopharyngealeareinoma CNE-2Z cells. Methods The eukaryotic expression vector of hsamiR-218 （pmiR-218） was constructed and transfected into CNE-2Z cells using LipofectamineTM 2000. The proliferation of CNE-2Z cells was examined by MTY essay. The cell apoptosis was analyzed using flow cytometry by double staining with Annexin V and Propidium Iodide. The level of Survivin protein was determined by Western blotting. Results The eukaryotic expression vector of hsa-miR- 218 （pmiR-218） was successfully constructed. The proliferation of CNE-2Z cells in thepmiR-218 group was inhibited compared with the negative control group （ P 〈 O. 05 ） . The apoptotic rates were much higher in the pmiR-218 group than in the negative control group （P 〈0. 05 ） . pmiR-218 downregulated the expression level of Survivin protein in CNE-2Z cells. Conclusion pmiR-218 results in inhibition of proliferation and induction of apoptosis in CNE-2Z ceils and the mechanism appears to be related with Survivin suppression by pmiR-218, indicating that hsa-miR-218 might be used as a potential therapeutic target in nasopharyngealeareinoma.