PIF1解旋酶在电离辐射诱发的细胞周期阻滞中的作用

发布时间：2018-07-06 16:25

本文选题：PIF解旋酶 + 辐射　；参考：《军事医学》2017年03期

【摘要】：目的观察人PIF1解旋酶敲低对电离辐射致细胞生长及细胞周期阻滞的影响,以探讨人PIF1在电离辐射致DNA损伤应答中的作用。方法采用慢病毒系统建立PIF1敲低稳转细胞系,实时荧光定量PCR及Western印迹检测PIF1敲低效果。细胞计数法观察PIF1敲低对4 Gyγ射线照射下细胞增殖的影响,流式细胞术观察PIF1敲低对8 Gyγ射线照射下细胞周期阻滞的影响。结果建立了PIF1敲低的稳转细胞系,4 Gyγ射线照射后,PIF1敲低细胞系的生长受到明显抑制,在照后第2~4天几乎无任何增殖,直到第5天才开始缓慢增殖。8 Gyγ射线照射后,PIF1敲低细胞系的S期阻滞与G_2/M期阻滞相对于对照细胞发生明显延迟。结论 PIF1敲低后显著增加了细胞的辐射敏感性以及电离辐射所致的S期阻滞与G_2/M期阻滞延迟。
[Abstract]:Aim to investigate the effects of low levels of human PIF1 helicase on cell growth and cell cycle arrest induced by ionizing radiation, and to explore the role of human PIF1 in DNA damage response induced by ionizing radiation. Methods Lentivirus system was used to establish PIF1 knockdown stable cell line, and real-time fluorescent quantitative PCR and Western blotting were used to detect the effect of PIF1 knockdown. Cell count method was used to observe the effect of PIF1 knockout on cell proliferation induced by 4 Gy 纬 -ray irradiation and flow cytometry to observe the effect of PIF1 knockout on cell cycle arrest induced by 8 Gy 纬 -ray irradiation. Results the growth of PIF1 knockdown cell line was significantly inhibited after 4 Gy 纬 -ray irradiation, and almost no proliferation was observed on the 2nd day after irradiation. It was not until the 5th day that the slow proliferation of PIF1 knockout cell lines in S phase arrest and G _ 2 / M phase arrest was significantly delayed compared with that of the control cells. Conclusion PIF1 knockout can significantly increase the radiosensitivity of the cells and delay the S phase arrest induced by ionizing radiation and the GSP 2 / M phase block.
【作者单位】：石河子大学医学院;石河子大学生命科学学院;军事医学科学院放射与辐射医学研究所;
【基金】：国家自然科学基金资助项目(31470827,81472910,81560473,31270894)
【分类号】：R594.8

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