RRC ID 3822
Author Takasaki I, Takarada S, Fukuchi M, Yasuda M, Tsuda M, Tabuchi Y.
Title Identification of genetic networks involved in the cell growth arrest and differentiation of a rat astrocyte cell line RCG-12.
Journal J. Cell. Biochem.
Abstract The purpose of the present study is to establish and characterize a conditionally immortalized astrocyte cell line and to clarify the genetic networks responsible for the cell growth arrest and differentiation. A conditionally immortalized astrocyte cell line, RCG-12, was established by infecting primary cultured rat cortical glia cells with a temperature-sensitive simian virus 40 large T-antigen. At a permissive temperature of 33 degrees C, the large T-antigen was expressed and cells grew continuously. On the other hand, the down-regulation of T-antigen at a non-permissive temperature of 39 degrees C led to growth arrest and differentiation. The cells expressed astrocyte-expressed genes such as glial fibrillary acidic protein. Interestingly, the differentiated condition induced by the non-permissive temperature significantly elevated the expression levels of several astrocyte-expressed genes. To identify the detailed mechanisms by which non-permissive temperature-induced cell growth arrest and differentiation, we performed high-density oligonucleotide microarray analysis and found that 556 out of 15,923 probe sets were differentially expressed 2.0-fold. A computational gene network analysis revealed that a genetic network containing up-regulated genes such as RB, NOTCH1, and CDKN1A was associated with the cellular growth and proliferation, and that a genetic network containing down-regulated genes such as MYC, CCNB1, and IGF1 was associated with the cell cycle. The established cell line RCG-12 retains some characteristics of astrocytes and should provide an excellent model for studies of astrocyte biology. The present results will also provide a basis for understanding the detailed molecular mechanisms of the growth arrest and differentiation of astrocytes.
Volume 102(6)
Pages 1472-85
Published 2007-12-15
DOI 10.1002/jcb.21369
PMID 17440958
MeSH Animals Antigens, Polyomavirus Transforming / metabolism Astrocytes / cytology Astrocytes / metabolism* Astrocytes / physiology* Cell Count Cell Differentiation* Cell Line, Transformed Cell Proliferation Cell Transformation, Viral Cerebral Cortex / cytology Computational Biology / methods Cyclin-Dependent Kinase Inhibitor p21 / metabolism Embryo, Mammalian Gene Expression Regulation / physiology* Gene Regulatory Networks* Glial Fibrillary Acidic Protein / metabolism Immunohistochemistry Models, Genetic Oligonucleotide Array Sequence Analysis Rats Rats, Sprague-Dawley Temperature Transfection
IF 2.959
Times Cited 11
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY CELL BIOLOGY
Resource
DNA material pSVtsA58 ori (-) (RDB01126)