Document Detail

Preferential differentiation of P19 mouse embryonal carcinoma cells into smooth muscle cells. Use of retinoic acid and antisense against the central nervous system-specific POU transcription factor Brn-2.
MedLine Citation:
PMID:  8593698     Owner:  NLM     Status:  MEDLINE    
Investigation of the molecular mechanisms that control smooth muscle cell (SMC) development and differentiation is a prerequisite in understanding the regulatory mechanisms of physiological and pathological SMC-associated vascular processes. The pluripotent murine embryonal carcinoma P19 cell, whose developmental potential resembles that of early embryonic cells, can develop into cell types derived from the neuroectoderm, mesoderm, and endoderm. In the present study, we have shown a unique strategy to enhance SMC differentiation in P19 cells. Under chemical induction of high concentrations of retinoic acid (1 micromol/L), P19 cells showed optimum differentiation into SMCs. Because the P19 cells thus induced also showed differentiation into neuronal cells, a strategy to block neuronal lineage differentiation was developed using a stable transformant antisense RNA construct against Brn-2, a neuronal lineage-specific POU-domain transcription factor; thus, by specifically inhibiting neuronal differentiation, enhanced SMC differentiation by P19 cells was attained. SMC expression was confirmed by immunohistochemical staining, RNA analysis (RNase protection assay), and protein analysis (Western blot) using SMC-specific markers (eg, SM1 and calponin) and alpha-smooth muscle actin. Our results show that the pathway of SMC differentiation may provide an in vitro system useful in the investigation of SMC regulatory mechanisms (eg, transcriptional regulation) and in the further understanding of SMC development and differentiation.
T Suzuki; H S Kim; M Kurabayashi; H Hamada; H Fujii; M Aikawa; M Watanabe; N Watanabe; Y Sakomura; Y Yazaki; R Nagai
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Circulation research     Volume:  78     ISSN:  0009-7330     ISO Abbreviation:  Circ. Res.     Publication Date:  1996 Mar 
Date Detail:
Created Date:  1996-04-10     Completed Date:  1996-04-10     Revised Date:  2005-11-17    
Medline Journal Info:
Nlm Unique ID:  0047103     Medline TA:  Circ Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  395-404     Citation Subset:  IM    
Third Department of Internal Medicine, University of Tokyo, Japan.
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MeSH Terms
Amino Acid Sequence
Base Sequence
Carcinoma, Embryonal / metabolism,  pathology*
Cell Differentiation
Fluorescent Antibody Technique, Indirect
Gene Expression
Homeodomain Proteins
Molecular Sequence Data
Muscle, Smooth / cytology*,  metabolism
POU Domain Factors
RNA, Antisense / genetics*
Transcription Factors / genetics*
Tretinoin / pharmacology*
Tumor Cells, Cultured
Reg. No./Substance:
0/Homeodomain Proteins; 0/POU Domain Factors; 0/RNA, Antisense; 0/Transcription Factors; 0/transcription factor Brn-2; 302-79-4/Tretinoin

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