INCREASED P53 PROTEIN ASSOCIATED WITH AGING IN HUMAN-DIPLOID FIBROBLASTS
INCREASED P53 PROTEIN ASSOCIATED WITH AGING IN HUMAN-DIPLOID FIBROBLASTS
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DOI：
10.1006/excr.1995.1095
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发表时间：
1995-04-01
影响因子：
3.7
通讯作者：
LEHMAN, JM
中科院分区：
文献类型：
--
作者：
KULJU, KS;LEHMAN, JM
通过分析肿瘤抑制蛋白p53的表达，研究了人二倍体成纤维细胞IMR-90株中与细胞衰老相关的分子变化。在所有研究中，IMR-90培养物根据形态学、群体倍增率、DNA合成能力和已建立的衰老标记物的存在被表征为“年轻"或”接近衰老“。当p53被单克隆抗体免疫沉淀并通过Western免疫印迹分析检测时，在近衰老培养物中检测到每个细胞更多的蛋白质。使用PAb 1801（N-末端特异性）抗p53抗体测得p53蛋白增加10倍以上，而PAb 122（C-末端特异性）测得增加5倍。虽然近衰老的文化表现出较高水平的p53比年轻的细胞，这些文化具有相似的电荷和分子量p53亚型时，通过二维蛋白质印迹分析。当p53 RNA与总RNA相比时，随着年龄的增长，p53 RNA减少，但在每个细胞的基础上，p53 RNA升高。这些结果为衰老过程中p53的转录调控提供了证据，并支持了p53蛋白水平升高可能在细胞衰老中发挥作用的假设。(C)出版社：Academic Press
Molecular changes associated with cellular aging in a strain of human diploid fibroblasts, IMR-90, were addressed by analyzing the expression of the tumor suppressor protein, p53. In all studies, IMR-90 cultures were characterized as ''young'' or ''near-senescent'' based on morphology, rate of population doubling, capacity for DNA synthesis, and presence of established markers for senescence. When p53 was immunoprecipitated by monoclonal antibodies and detected by Western immunoblot analysis, more protein per cell was detected in the near-senescent cultures. A greater than 10-fold increase in p53 protein was measured with the PAb 1801 (N-terminal-specific) anti-p53 antibody, whereas PAb 122 (C-terminal-specific) measured a 5-fold increase. Although near-senescent cultures demonstrated a higher level of p53 than young cells, these cultures had similar charges and molecular weight p53 isoforms when analyzed by two-dimensional Western blots. When p53 RNA was compared to total RNA there was a decrease in p53 RNA with age, but on a per cell basis p53 RNA was elevated. These results provide evidence for transcriptional regulation of p53 during aging and support the hypothesis that elevated levels of p53 protein may play a role in cellular senescence. (C) 1995 Academic Press, Inc.