[Accelerated somatic cell senescence and changes in p16INK4a expression after exogenous DNA transfection]

ArticleinHereditas (Beijing) 28(11):1383-8 · December 2006with1 Reads
DOI: 10.1360/yc-006-1383 · Source: PubMed
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.
  • [Show abstract] [Hide abstract] ABSTRACT: The fatty acid dehydrogenase I (FatI) is able to express in mammalian cells and convert n-6 PUFAs to n-3 PUFAs. n-3 polyunsaturated fatty acids (n-3 PUFAs) is an important component of cell membrane, plays an important role in prevention and control variety of human disease. However, n-3 PUFAs cannot be endogenously synthesized by mammals because they lack the dehydrogenase converting n-6 to n-3 PUFA. As time being, gradually matured transgenic technology makes it possible to produce transgenic animals that are able to synthesize n-3 PUFAs by themselves. However, the transgenic technology itself may bring negative impacts. In this study, the eukaryotic expression vector pcDNA3.1-FatI was introduced into genome of Boer goat fetal fibroblasts cultured in vitro, and the influence of biological characteristics of fetal fibroblast was studied via overexpression of FatI. The results showed the proliferation and apoptosis of cultured fetal fibroblast were not affected significantly by the overexpression of FatI using BrdU and TUNEL staining methods, respectively. Moreover, the overexpression of FatI significantly inhibited the senescence of somatic cells compared to EGFP transgenic cells (p < 0.01). Quantitative PCR revealed the mRNA expression of P16 and P53 in the FatI transgenic cells group were significantly lower than that of EGFP transgenic cells group (p < 0.01). In conclusion, the senescence of goat somatic cells was inhibited by the over-expression of FatI gene. This article is protected by copyright. All rights reserved
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