shRNA干扰沉默Net1基因对电离辐射损伤反应的影响

The effects of short hairpin RNA-mediated silencing Net1 on ionizing radiation-induced damage responses

    摘要
  • 摘要:
    目的 研究神经上皮细胞转化基因1（NET1）在电离辐射损伤反应中的生物学功能。
    方法 运用RNA干扰技术，在细胞中特异性沉默Net1基因，然后通过集落存活实验和免疫印迹法研究抑制Net1表达对细胞的电离辐射损伤反应的影响。
    结果 Net1敲低表达的细胞对电离辐射的敏感性增强，表现为更多细胞发生了凋亡，并且在γ射线照射后，毛细血管扩张性共济失调症突变蛋白（ATM）和细胞周期检查点激酶2（Chk2）的磷酸化水平也比对照细胞增强。
    结论 Net1对于受到电离辐射的细胞有一定的保护作用，很可能在电离辐射损伤修复中具有重要作用。

     

    Abstract:
    Objective To study the biological roles of the neuroepithelioma transforming gene 1 (Net1)in the cellular responses to ionizing radiation(IR)-induced damage.
    Methods Specific shRNA was used to deplete Net1 in cells. The effects of Net1 depletion on the cellular responses to ionizing radiation were investigated through the colonogenic survival assay and immunoblotting analysis of DNA damage response proteins′ phosphorylation.
    Results Net1-depleted cells were more sensitive to IR, with a significantly increased induction of apoptosis. In response to IR, the phosphorylation levels of ataxia-telangiectasia mutated and checkpoint kinase 2 were much higher in the Net1-depleted cells than that in the control cells.
    Conclusion Net1 protects IR-treated cells from apoptosis and possibly plays an important role in IR-induced damage response and repair.

     

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