EVALUATION OF CELL-DEATH IN EBV-TRANSFORMED LYMPHOCYTES USING AGAROSE-GEL ELECTROPHORESIS, LIGHT-MICROSCOPY AND ELECTRON-MICROSCOPY .2. INDUCTION OF NONCLASSIC APOPTOSIS (PARA-APOPTOSIS) BY TRITIATED-THYMIDINE
EVALUATION OF CELL-DEATH IN EBV-TRANSFORMED LYMPHOCYTES USING AGAROSE-GEL ELECTROPHORESIS, LIGHT-MICROSCOPY AND ELECTRON-MICROSCOPY .2. INDUCTION OF NONCLASSIC APOPTOSIS (PARA-APOPTOSIS) BY TRITIATED-THYMIDINE
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DOI：
10.3109/10428199509059664
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发表时间：
1995-09-01
影响因子：
2.6
通讯作者：
BERNSTEIN, C
中科院分区：
文献类型：
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作者：
ASHER, E;PAYNE, CM;BERNSTEIN, C
有大量的文献可以追溯到20世纪50年代后期，关于放射性标记DNA在体内的破坏性生物效应。尽管如此，氚标记的胸苷在程序性细胞死亡（凋亡）的研究中经常被用来标记DNA。在本研究中，我们研究了在没有任何其他凋亡诱导剂的情况下，将氚化胸苷掺入EB病毒转化细胞系（NC-37）的DNA中的影响。将细胞在含有1-20 μ Ci/ml [甲基-H-3]-胸苷（[H-3]-TdR）的培养基中孵育。在使用的每个浓度的氚化胸苷下，细胞增殖在孵育12小时内停止。使用DNA降解模式和细胞形态学评价氚化胸苷掺入引起的细胞死亡模式。琼脂糖凝胶电泳显示DNA降解，无“梯状"模式。电子显微镜被用作“金标准”，以评估伴随DNA降解的细胞死亡的特定形态学类型。虽然存在细胞凋亡的一些特征，但细胞缺乏完整核内染色质的早期边缘化和导致凋亡小体形成的表面起泡，这是细胞凋亡的两个特征性形态学特征。因此，我们创造了术语“准凋亡”，以更精确地描述细胞死亡的形态学类型。准凋亡细胞的百分比通过光学显微镜使用整体封片制备物（细胞离心涂片）进行定量。染色质凝聚，核碎裂，细胞密度增加和细胞质空泡化的形态学标准被用于通过光显微镜的细胞离心涂片制备的准凋亡的评价。在没有氚化胸苷的情况下，
There is an extensive literature dating back to the late 1950's, on the damaging biological effects of radiolabeling DNA in vivo. Nonetheless, tritiated thymidine has often been used to label DNA in studies of programmed cell death (apoptosis). In the present study, we have investigated the effects of incorporation of tritiated thymidine into the DNA of an Epstein-Barr virus-transformed cell line (NC-37) in the absence of any other apoptosis-inducing agent. Cells were incubated in media containing 1-20 mu Ci/ml [methyl-H-3]-thymidine ([H-3]-TdR). At each concentration of tritiated thymidine used, cell proliferation ceased within 12 hours of incubation. The mode of cell death caused by tritiated thymidine incorporation was evaluated using DNA degradation patterns and cellular morphology. DNA degradation, in the absence of a ''ladder'' pattern, was shown by agarose gel electrophoresis. Electron microscopy was used as the ''gold standard'' to evaluate the specific morphologic type of cell death that accompanied the DNA degradation. Although some of the features of apoptosis were present, the cells lacked the early margination of the chromatin within an intact nucleus and surface blebbing leading to apoptotic body formation, two characteristic morphological features of apoptosis. We, therefore, coined the term ''para-apoptosis'' to be more precise about the morphologic type of cell death, The percent of para-apoptotic cells was quantitated by light microscopy using whole mount preparations (cytospins). The morphologic criteria of chromatin condensation, nuclear fragmentation, increase in cell density and cytoplasmic vacuolization were used for the evaluation of para-apoptosis by light microscopy of cytospin preparations. In the absence of tritiated thymidine,