论文部分内容阅读
目的 定性和定量地分析放射诱导ＨＬ６０ 细胞凋亡。 方法 以人类白血病细胞系ＨＬ６０ 为材料，应用光学显微镜和电子显微镜及琼脂糖电泳定性分析６０ Ｃｏ 放射线诱导ＨＬ６０ 细胞凋亡的发生，应用ＤＮＡ流式细胞计量术和末端转移酶（ＴｄＴ） 介导的缺口末端标记法（ＴＵＮＥＬ） 的流式细胞定量检测ＨＬ６０ 细胞凋亡。 结果 （１） 放射后ＨＬ６０ 出现细胞体积缩小，染色质固缩致密化、边缘化，染色体断裂，凋亡小体和细胞膜保持完整等形态学特征。（２） 琼脂糖电泳表现为特征性ＤＮＡ“梯谱”。（３）流式细胞计量术发现放射诱导ＨＬ６０ 细胞凋亡在照射后６ｈ 已较明显，而且细胞凋亡百分比随着照射剂量增加而增加。ＴＵＮＥＬ法检测细胞凋亡百分比值比ＤＮＡ流式细胞分析的方法得到的值要小，但差异无显著意义。 结论 流式细胞计量术结合细胞形态学检查和（ 或） 琼脂糖电泳能较好、简便、迅速地分析放射诱导ＨＬ６０ 细胞凋亡
Objective To qualitatively and quantitatively analyze the radiation-induced apoptosis of HL-60 cells. METHODS: Human leukemia cell line HL60 was used as material. The apoptosis of HL60 cells was induced by 60 Co radiation using light microscopy, electron microscopy, and agarose electrophoresis. DNA flow cytometry and terminal transferase (TdT) were used. Apoptosis was detected by TUNEL-mediated flow cytometry in HL-60 cells. Results (1) After irradiation, HL-60 appeared to have a morphological feature such as a decrease in cell volume, chromatin condensation and densification, marginalization, chromosomal breakage, apoptotic bodies and cell membrane integrity. (2) Agarose electrophoresis appears as a characteristic DNA “ladder”. (3) Flow cytometry found that radiation-induced apoptosis of HL-60 cells was evident at 6 h after irradiation, and the percentage of apoptosis increased with increasing irradiation dose. The percentage of apoptosis detected by TUNEL method was smaller than that obtained by DNA flow cytometry analysis, but the difference was not significant. Conclusion Flow cytometry combined with cell morphology and (or) agarose electrophoresis can be used to analyze radiation-induced apoptosis of HL-60 cells in a good, simple and rapid manner.