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低氧诱导U251细胞中miRNA-210的表达及对肿瘤转移的影响

Expression of miRNA-210 Induced by Hypoxia in U251 Cells and Effect on Metastasis

  • 摘要: 目的 研究低氧条件下胶质瘤细胞系U251细胞中miRNA-210的表达以及对肿瘤转移能力的影响。方法 低氧条件(1% O2)培养胶质瘤细胞系U251,利用Western blot检测不同诱导时间HIF-1α水平变化, 利用 real time PCR检测低氧不同诱导时间miRNA-210表达变化,通过Transwell小室检测U251 细胞侵袭能力,利用Western blot检测低氧条件下Vmp1表达变化。结果 低氧诱导U251细胞6、12和24h,可检测到HIF-1α水平随着诱导时间而增加;在低氧诱导的不同时间点可检测到miRNA-210表达逐渐增加,给予HIF-1α抑制剂2-甲氧雌二醇(2-methoxyestradiol,2-ME2)后miRNA-210的表达受到抑制;低氧条件下U251细胞侵袭转移能力增强,在miRNA-210抑制剂antagonist miR210转染后,U251细胞转移能力受到抑制,低氧条件下U251细胞中Vmp1蛋白表达下降,antagonist miR210转染可以逆转Vmp1的表达。结论 低氧可以通过HIF-1α诱导miRNA-210表达增加,miRNA-210具有促进U251细胞转移的作用,miRNA-210可能通过调节Vmp1表达发挥作用。

     

    Abstract: Objective To investigate the effects of hypoxia on expression of miRNA-210 and U251 cell metastasis. Methods Human glioma U251 cell line was induced by hypoxia(1% O2) for different hours. The level of Hypoxia-inducible factor 1α(HIF-1α) in U251 cells was detected by western blot and the expression of miRNA-210 was tested by real time PCR. The effects of miRNA-210 on U251 cells metastasis were tested by tumor invasion assay using Transwell. The expression of Vmp1 was tested by western blot. Results The level of HIF-1α in U251 cells were signifi cantly increased in time-dependent manner when induced for 6h, 12 h, and 24 h by hypoxia, and the expression of miRNA-210 was increased with HIF-1α level. The expression of miRNA-210 was suppressed by 2-ME2, HIF-1α inhibitor. The metastasis capability of U251 cells induced by hypoxia was increased, and was inhibited by miRNA210 inhibitor antagonist miR210. The expression of Vmp1 in U251 cells induced by hypoxia was increased and reversed by antagonist miR210. Conclusion miRNA-210 could be induced by hypoxia via HIF-1α. miRNA-210 could promote U251 cells metastasis and down-regulate Vmp1 expression .

     

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