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《Acta Zoologica Sinica》 2002-01
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CHANGES IN KERATIN DURING APOPTOSIS OF PK15 (PORCINE KIDNEY-15) CELLS

CHEN Dan-Ying GAO Yun-Fei LI Li-Xia ZHAI Zhong-He (Life Science College, Peking University, Beijing 100871, China)  
Intermediate filament (IF) is a major component of cytoskeleton.Recent advances suggest that IFs not only play a cytoskeletal role in cooperation with microfilaments and microtubules, but are also involved in cell differentiation, signal transduction and gene expression.Apoptosis is a physiological form of cell death, it describes the orchestrated collapse of a cell, staging membrane blebbing, cell shrinkage, protein fragmentation, chromatin condensation and DNA degradation.How do IFs change in apoptotic process and what kind of role do they play? The answers to these questions would be of great importance to further study of their functions.Epithelial cell PK15(Porcine Kidney-15) was induced to apoptosis by actinomycin D.The cell shapes changed from spreading to contracting and rounding during the induction process, and most of the cells dropped from the wall after induction for 96 h.DAPI staining showed the chromatin condensation and apoptotic bodies formation characterized by apoptosis in this process.At the same time “DNA ladder” was detected by electrophoresis.Indirect immunofluorescence results showed that the fluorescence of keratin distributed unevenly in the apoptotic cells, and in some cells the keratin fluorescence contracted around the condensed chromatin.Using selective extraction combined with whole mount electron microscopy, we detected the intact keratin filament network in some area of apoptotic cells, and the filaments were obviously shortened.In consistent with the result of immunofluorescence, the keratin filaments around chromatin were condensed in comparison with filaments in other areas.Western blot analysis further showed that part of type Ⅱ keratin protein degraded only partially during apoptosis, whereas type Ⅰ keratin protein molecules were intact.These results suggested that in apoptotic processing the structure changes of keratin network might lead to the changes of cell shape, and might participate the formation of apoptotic bodies.
【Fund】: 国家重点基础研究发展规划项目 (No G19990 5 39)~~
【CateGory Index】: Q255
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