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目的研究X射线辐照后GC1小鼠精原细胞凋亡诱导因子(AIF)细胞定位的变化。方法 GC1细胞分别给予0、3、6、9 Gy X射线辐照,溴脱氧尿苷(Brd U)掺入实验检测辐照对细胞增殖速率的影响;辐照后48 h,4',6-二脒基-2-苯基吲哚(DAPI)染色观察细胞核凝集情况,免疫荧光细胞化学染色检测胞质及胞核中AIF蛋白定位的变化,Western blot法分析细胞总蛋白、胞质蛋白及核提取物中AIF蛋白的水平。结果随着辐照剂量的增加,GC1细胞增殖速率显著减慢,细胞活性降低。6 Gy X射线辐照后全细胞提取物中AIF蛋白总量变化不大,但细胞核提取物中AIF显著增多,即AIF出现核转位的现象。结论 X射线辐照能够诱导小鼠精原细胞发生凋亡,该过程中伴有AIF入核的现象。
Objective To study the changes of the localization of apoptosis-inducing factor (AIF) cells in GC1 mice after X-ray irradiation. Methods GC1 cells were irradiated with 0, 3, 6, and 9 Gy X-rays, respectively. BrdU incorporation assay was used to detect the effect of irradiation on cell proliferation rate. At 48 h after irradiation, DAPI staining was used to observe the nuclear agglutination. Immunofluorescence staining was used to detect the localization of AIF protein in the cytoplasm and nucleus. The total protein, cytoplasmic protein and nucleus were analyzed by Western blot AIF protein levels in the extract. Results With the increase of radiation dose, the proliferation rate of GC1 cells decreased significantly and the cell activity decreased. The total amount of AIF protein in whole cell extracts after 6 Gy X-ray irradiation did not change much, but AIF in nuclear extracts was significantly increased, that is, AIF appeared nuclear translocation. Conclusion X-ray irradiation can induce the apoptosis of mouse spermatogonia, which is accompanied by the phenomenon of AIF entry into the nucleus.