Martelli A M, Bortul R, Fackelmayer F O, Tazzari P L, Bareggi R, Narducci P, Zweyer M
Dipartimento di Morfologia Umana Normale, Università di Trieste, Italy.
J Cell Biochem. 1999 Jan 1;72(1):35-46. doi: 10.1002/(sici)1097-4644(19990101)72:1<35::aid-jcb5>3.0.co;2-s.
We have characterized the nuclear matrix-intermediate filament fraction from control and apoptotic HL-60 cells. Apoptosis was induced by exposure to the topoisomerase I inhibitor, camptothecin. By means of two-dimensional polyacrylamide gel electrophoresis, striking qualitative and quantitative differences were seen in the protein composition of the nuclear matrix-intermediate filament fraction obtained from apoptotic cells in comparison with controls. Western blotting analysis of apoptotic nuclear matrix proteins revealed degradation of some (topoisomerase IIalpha, SAF-A) but not other (SATB1 and nucleolin) components. Moreover, immunofluorescent staining for typical matrix antigens (NuMA protein, lamin B, SC-35) showed that in 35-40% of the structures prepared from apoptotic samples, marked changes in the subnuclear distribution of these proteins were present. Striking morphological differences between control and apoptotic samples were also detected at the ultrastructural level. These results demonstrate that both biochemical and morphological changes can be detected in the nuclear matrix prepared from apoptotic HL-60 cells.
我们已对来自对照和凋亡HL-60细胞的核基质-中间丝组分进行了表征。通过暴露于拓扑异构酶I抑制剂喜树碱来诱导细胞凋亡。借助二维聚丙烯酰胺凝胶电泳，与对照相比，在从凋亡细胞获得的核基质-中间丝组分的蛋白质组成中观察到了显著的定性和定量差异。对凋亡核基质蛋白的蛋白质免疫印迹分析显示，一些组分（拓扑异构酶IIα、SAF-A）发生了降解，但其他组分（SATB1和核仁素）未发生降解。此外，对典型基质抗原（NuMA蛋白、核纤层蛋白B、SC-35）的免疫荧光染色显示，在从凋亡样品制备的结构中，有35%-40%的结构中这些蛋白质的核内分布存在明显变化。在超微结构水平上也检测到了对照样品和凋亡样品之间显著的形态学差异。这些结果表明，在从凋亡HL-60细胞制备的核基质中可以检测到生化和形态学变化。
