X射线辐照引起GC1小鼠精原细胞凋亡诱导因子核转位并诱导其凋亡

发布时间：2018-10-20 20:47

【摘要】：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。
[Abstract]:Objective to study the localization of spermatogonial apoptosis-inducing factor (AIF) in GC1 mice after X-ray irradiation. Methods GC1 cells were irradiated with X ray for 0 ~ 3X ~ (69) Gy, respectively. The effect of irradiation on cell proliferation was detected by bromodeoxyuridine (Brd U) incorporation assay, and the nuclear agglutination was observed by 48 h ~ (48 h) after irradiation with 6-diamidine-2-phenylindole (DAPI) staining. The changes of AIF protein localization in cytoplasm and nucleus were detected by immunofluorescence cytochemical staining. The levels of AIF protein in total cellular protein, cytoplasmic protein and nuclear extract were analyzed by, Western blot method. Results with the increase of irradiation dose, the proliferation rate of GC1 cells decreased significantly, and the cell activity decreased. The total AIF protein content in the whole cell extract was not changed after X ray irradiation for 6 Gy, but the AIF in the nuclear extract increased significantly. This is the phenomenon of nuclear translocation in AIF. Conclusion X-ray irradiation can induce apoptosis of spermatogonial cells in mice, which is accompanied by the phenomenon of AIF entry into the nucleus.
【作者单位】：宁夏医科大学生育力保持教育部重点实验室;宁夏医科大学生物化学与分子生物学系;宁夏医科大学总医院肿瘤医院;宁夏医科大学生理学系;
【基金】：国家自然科学基金(81460478) 宁夏教育厅优青项目(NGY2015095)
【分类号】：R818

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