遗传 ›› 2006, Vol. 28 ›› Issue (11): 1383-1388.doi: 10.1360/yc-006-1383

• 研究报告 • 上一篇    下一篇

外源基因转染导致小鼠体细胞过快衰老 及p16表达水平变化

沈 伟1~3; 李 兰1,3; 吴晓洁1; 周艳荣1; 潘庆杰3; 陈 宏2; 邓继先1   

  1. 1. 军事医学科学院生物工程研究所, 北京 100071; 2. 西北农林科技大学动物科技学院, 杨凌 712100;
    3. 莱阳农学院动物科技学院动物生殖发育与基因工程研究所, 青岛 266109

  • 收稿日期:2005-12-20 修回日期:2006-01-12 出版日期:2006-11-05 发布日期:2006-11-05
  • 通讯作者: 沈伟

Accelerated Somatic Cell Senescence and Changes in p16 Expression after Exogenous DNA Transfection

SHEN Wei1~3; LI Lan1,3; WU Xiao-Jie1; ZHOU Yan-Rong1; PAN Qing-Jie3; CHEN Hong2;
DENG Ji-Xian1

  

  1. 1. Institute of Biotechnology, Academy of Military Medical Science, Beijing 100071, China; 2. Department of Animal Sciences, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100, China; 3. Institute of Animal Reproduction, Development and Genetic Engineering, Department of Animal Sciences, Laiyang Agriculture College, Qingdao 266109, China
  • Received:2005-12-20 Revised:2006-01-12 Online:2006-11-05 Published:2006-11-05
  • Contact: SHEN Wei

摘要:

对小鼠胎儿成纤维细胞进行外源基因转染时发现, 外源基因转染后的小鼠体细胞染色体端粒的长度以每代47 bp碱基缩短; 在转染后的衰老细胞中, 或细胞随着增龄, p16INK4a 5′-调控区DNA甲基化程度逐渐降低; 利用RT-PCR与Northern blot证明, 衰老细胞与年轻细胞中的p16INK4a基因的表达水平存在显著差异, 传代45代的细胞和外源基因转染后的衰老细胞p16INK4a基因的表达水平大约是原代细胞的12~16倍, 而原代细胞与20代细胞间的差异很小。外源基因转染后的衰老细胞核移植后能支持克隆胚胎的体外早期发育。

关键词: p16INK4a基因, 端区, 细胞衰老, 转基因, 小鼠

Abstract:

During the transfection of mouse embryonic fibroblasts (MEF), we found these cells became senescent, appearing enlarged with hollow cytoplasm and multinucleated. The telomere lengths of these senescent MEFs were significantly shorter than primary untransfected MEFs. In senescent MEF cells, DNA methylation of p16INK4a 5’-regulatory region showed gradual reduction as cells aged. RT-PCR and Northern blot demonstrated that the expression of p16INK4a in transfected senescent cells was 12-16 times more than primary cells. The senescent transfected MEF cells as donors of nuclei could support the early development of cloned embryos after nuclear transfer.

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