Huschtscha L I, Jeitner T M, Andersson C E, Bartier W A, Tattersall M H
Department of Cancer Medicine, University of Sydney, New South Wales, Australia.
Exp Cell Res. 1994 May;212(1):161-5. doi: 10.1006/excr.1994.1131.
Glucocorticoids kill human leukemic cells, CCRF-CEM.f2, by apoptosis. Cell death is preceded by DNA fragmentation into nucleosomal subunits which can be evaluated by DNA gel electrophoresis or by determining the proportion of cleaved DNA in whole cell lysates. Hyperthermic treatment of CCRF-CEM.f2 cells induces necrotic cell death which is characterized by a different DNA gel electrophoretic pattern. However, these techniques cannot distinguish the individual contributions of both forms of death in a heterogeneous population. Therefore, we have developed a flow cytometric method that readily distinguishes apoptotic and necrotic cells on the basis of propidium iodide staining and cellular light scatter characteristics. This method can be used to analyze factors influencing the mechanisms of cell death after cytotoxic drug treatment.
糖皮质激素通过凋亡杀死人白血病细胞CCRF-CEM.f2。细胞死亡之前会发生DNA断裂成核小体亚基的现象，这可以通过DNA凝胶电泳或通过测定全细胞裂解物中切割DNA的比例来评估。对CCRF-CEM.f2细胞进行高温处理会诱导坏死性细胞死亡，其特征是具有不同的DNA凝胶电泳图谱。然而，这些技术无法区分异质群体中两种死亡形式各自的作用。因此，我们开发了一种流式细胞术方法，该方法基于碘化丙啶染色和细胞光散射特性能够轻松区分凋亡细胞和坏死细胞。该方法可用于分析影响细胞毒性药物治疗后细胞死亡机制的因素。
