沉默BLAP75基因对电离辐射诱导DNA损伤的影响

Effects of silencing BLAP75 on DNA damage induced by ionizing radiation

    摘要
  • 摘要:
    目的 研究BLM结合蛋白75（BLAP75）在电离辐射诱导DNA损伤反应中的生物学效应。
    方法 运用RNA干扰技术，在细胞中特异性沉默BLAP75基因，然后通过单细胞凝胶电泳技术来研究电离辐射诱导DNA损伤程度的变化，并且通过再次表达BLAP75来拯救沉默BLAP75所致的实验表型，以及应用Western blot方法研究DNA损伤反应中的磷酸化修饰。
    结果 与未转染的293T对照组细胞相比，电离辐射在沉默了BLAP75基因的细胞中会诱导更多的DNA断裂，并且在此细胞中重新表达BLAP75则能降低DNA断裂数量至对照组细胞水平；γ射线照射后，细胞周期检查点激酶2（Chk2）的磷酸化程度比阴性对照组细胞增强。
    结论 BLAP75能减少电离辐射诱导的DNA损伤，在电离辐射损伤修复中可能具有重要作用。

     

    Abstract:
    Objective To study the biological effects of BLM-associated protein 75(BLAP75) in ionizing radiation(IR) induced DNA damage response(DDR).
    Methods BLAP75 was silenced in cells by RNA interference. Single-cell gel electrophoresis was performed to quantify the DNA breaks. Also siRNA-resistant BLAP75 was expressed in BLAP75-silenced cells to rescue the phenotype. Western blot was used to examine the DDR phosphorylation upon IR.
    Results IR induced more DNA breaks in BLAP75-silenced cells than in 293T control cells without siRNA transfection. After expression of BLAP75 in BLAP75-silenced cells, IR induced DNA breaks were similar to 293T control cells. In response to IR, the phosphorylation level of Chk2 was higher in the BLAP75-silenced cells than in the control cells.
    Conclusion BLAP75 alleviates IR-induced DNA damage and possibly plays an important role in IR-induced damage response and repair.

     

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