Document Detail


In vitro transformation of mouse testis cells by oncogene transfection.
MedLine Citation:
PMID:  22357549     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Germ cell tumors (GCTs) are unique in that they exhibit diverse biological characteristics and pathological features. Although several in vivo GCT models are available, studies on GCTs are hampered because in vivo development of GCTs is time consuming and prevents a detailed molecular analysis of the transformation process. Here we developed a novel strategy to transform mouse testis cells in vitro. Lentivirus-mediated transfection of dominant negative Trp53, Myc, and activated Hras1 into a CD9-expressing testis cells caused tumorigenic conversion in vitro. Although these cells resembled embryonic stem (ES) cells, they were aneuploid and lacked Nanog expression, which is involved in the maintenance of the undifferentiated state in ES cells. Euploid ES-like cells were produced by transfecting the Yamanaka factors (Pou5f1, Myc, Klf4, and Sox2) into the same cell population. Although these cells expressed Nanog, they were distinct from ES cells in that they expressed CD44, a cancer stem cell antigen. Both treatments induced similar changes in the DNA methylation patterns in differentially methylated regions of imprinted genes. Moreover, despite the differences in their phenotype and karyotype, both cell types similarly produced mixed GCTs on transplantation, which were composed of teratomas, seminomas, and embryonal carcinomas. Thus, in vitro testis cell transformation facilitates an analysis of the GCT formation process, and our results also suggest the close similarity between GCT formation and reprogramming.
Authors:
Hiroko Morimoto; Jiyoung Lee; Takashi Tanaka; Kei Ishii; Shinya Toyokuni; Mito Kanatsu-Shinohara; Takashi Shinohara
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-05-10
Journal Detail:
Title:  Biology of reproduction     Volume:  86     ISSN:  1529-7268     ISO Abbreviation:  Biol. Reprod.     Publication Date:  2012 May 
Date Detail:
Created Date:  2012-05-11     Completed Date:  2012-09-17     Revised Date:  2012-12-06    
Medline Journal Info:
Nlm Unique ID:  0207224     Medline TA:  Biol Reprod     Country:  United States    
Other Details:
Languages:  eng     Pagination:  148, 1-11     Citation Subset:  IM    
Affiliation:
Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
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MeSH Terms
Descriptor/Qualifier:
Aneuploidy
Animals
Antigens, CD44 / analysis
Antigens, CD9 / analysis
Carcinoma, Embryonal / genetics
Cell Transformation, Neoplastic / genetics*
DNA Methylation
Genomic Imprinting
Homeodomain Proteins / analysis
Kruppel-Like Transcription Factors / genetics
Lentivirus Infections
Male
Mice
Neoplasms, Germ Cell and Embryonal / genetics*
Octamer Transcription Factor-3 / genetics
Oncogenes*
Proto-Oncogene Proteins c-myc / genetics
Proto-Oncogene Proteins p21(ras) / genetics
SOXB1 Transcription Factors / genetics
Seminoma / genetics
Teratoma / genetics
Testicular Neoplasms / genetics*
Testis*
Transfection / methods*
Chemical
Reg. No./Substance:
0/Antigens, CD44; 0/Antigens, CD9; 0/Cd44 protein, mouse; 0/Cd9 protein, mouse; 0/GKLF protein; 0/Homeodomain Proteins; 0/Kruppel-Like Transcription Factors; 0/Nanog protein, mouse; 0/Octamer Transcription Factor-3; 0/Pou5f1 protein, mouse; 0/Proto-Oncogene Proteins c-myc; 0/SOXB1 Transcription Factors; 0/Sox2 protein, mouse; EC 3.6.5.2/Kras2 protein, mouse; EC 3.6.5.2/Proto-Oncogene Proteins p21(ras)

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