Document Detail


Phenotypic conversion of SV40-immortalized human diploid fibroblasts to senescing cells by introduction of an antisense gene for SV40-T antigen.
MedLine Citation:
PMID:  1338304     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Normal human lung fibroblast diploid cells, WI-38, become senescent after a definite number of divisions. VA-13 is a line of immortalized cells established by transformation of WI-38 cells by SV40 virus. To determine whether SV40 large T (SV40-T) antigen is essential for this immortalization of WI-38 cells we introduced an antisense gene for T antigen into VA-13. Two morphologically different types of antisense transformant (VA-AS5-8 and VA-AS37-8) were obtained. In both antisense transformants the expression of T antigen was reduced by more than 70% as compared to that in the parent cells. The morphology of the antisense transformants indicated a partial conversion to the senescent phenotype of WI-38. The relative number of cells in the S phase of the antisense transformants was decreased as compared to that in cultures of VA-13 and about 50% of cells were at G1/0. The doubling time of the transformants was prolonged to close to the doubling time of WI-38. The level of expression of retinoblastoma protein (pRB) complexed with SV40-T antigen of the antisense transformants was significantly decreased although the level of total pRB was much higher than that in VA-13. The pRB was present exclusively in the underphosphorylated form. Thus, the decreased level of formation of the complex between SV40-T and pRB or the underphosphorylation of pRB may explain the suppression of growth of antisense transformants. Together, these results show that an antisense gene for SV40-T antigen can efficiently block the cell proliferation and the cell immortalization of VA-13 cells.
Authors:
Y Tanaka; X Tang; D X Hou; H Gao; I Kitabayashi; G Gachelin; K Yokoyama
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cell structure and function     Volume:  17     ISSN:  0386-7196     ISO Abbreviation:  Cell Struct. Funct.     Publication Date:  1992 Dec 
Date Detail:
Created Date:  1993-04-20     Completed Date:  1993-04-20     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7608465     Medline TA:  Cell Struct Funct     Country:  JAPAN    
Other Details:
Languages:  eng     Pagination:  351-62     Citation Subset:  IM    
Affiliation:
Gene Bank, Tsukuba Life Science Center, RIKEN (The Institute of Physical and Chemical Research, Ibaraki, Japan.
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MeSH Terms
Descriptor/Qualifier:
Antigens, Polyomavirus Transforming / genetics*
Cell Aging*
Cell Cycle
Cell Transformation, Viral / genetics*
DNA, Antisense / genetics*
Fibroblasts / pathology*
Gene Expression Regulation, Viral
Humans
Phenotype
Phosphorylation
Retinoblastoma Protein / biosynthesis
Simian virus 40 / genetics,  physiology*
Chemical
Reg. No./Substance:
0/Antigens, Polyomavirus Transforming; 0/DNA, Antisense; 0/Retinoblastoma Protein

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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