Alterations of Expression of the Cytoskeleton after Immortalization of Human Fibroblasts

Cell Structure and Function

Yuji Karasaki²  Shigeru Kaneko²  Taro Ohji²  Ken Higashi²  Yasuhiro Satoh²  Sadao Gotoh²  Tetsumi Konishi²  Kunio Ikemura¹ 

[1] Department of Oral Surgery, University of Occupational and Environmental Health;Department of Biochemistry, University of Occupational and Environmental Health

关键词: cytoskeleton;    human fibroblast;    senescence;    immortalization;   

DOI  :  10.1247/csf.20.107

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(53)Cited-By(10)Human diploid fibroblasts (HDF) have a limited in vitro lifespan of population doubling level (PDL) 50-70. The molecular mechanism underlying cellular senescence and immortalization is not thoroughly understood. It has been reported that the cytoskeleton has diverse functions and may have a role in growth regulation through association with other cellular components. To shed light on the relationship between functions of the cytoskeleton and senescence or immortalization, We investigated the alterations in gene expression after immortalization and measured the amounts of mRNAs for talin, vinculin, α-actinin, tropomyosin 1 (TM1), vimentin, lamin A and C, and α-tubulin by slot blot and Northern blot analyses. We found that the mRNAs for vinculin and vimentin were reduced and the mRNA for lamin A was increased in immortalized cells. We also studied the cytoskeletal protein levels and their intracellular distributions by Western blot analysis and immunostaining. Most of the proteins studied behaved in a way similar to the mRNAs through senescence and immortalization. Vinculin, tropomyosins and vimentin showed their altered distributions in immortalized cells.

【 授权许可】

Unknown