title: In vitro transformation of mouse testis cells by oncogene transfection.
abstract: 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.affiliation: Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Morimoto
authors.forename: Jiyoung
authors.initials: J
authors.lastname: Lee
authors.forename: Takashi
authors.initials: T
authors.lastname: Tanaka
authors.forename: Kei
authors.initials: K
authors.lastname: Ishii
authors.forename: Shinya
authors.initials: S
authors.lastname: Toyokuni
authors.forename: Mito
authors.initials: M
authors.lastname: Kanatsu-Shinohara
authors.forename: Takashi
authors.initials: T
authors.lastname: Shinohara
mesh_terms: D000782:Aneuploidy; D000818:Animals; D018236:Carcinoma, Embryonal / Q000235:genetics; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D019175:DNA Methylation; D018392:Genomic Imprinting; D018398:Homeodomain Proteins / Q000032:analysis; D018960:Hyaluronan Receptors / Q000032:analysis; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics; D016180:Lentivirus Infections; D008297:Male; D051379:Mice; D000071317:Nanog Homeobox Protein; D009373:Neoplasms, Germ Cell and Embryonal / Q000235:genetics*; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D009857:Oncogenes; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000235:genetics; D018239:Seminoma / Q000235:genetics; D013724:Teratoma / Q000235:genetics; D013736:Testicular Neoplasms / Q000235:genetics*; D013737:Testis; D060245:Tetraspanin 29 / Q000032:analysis; D014162:Transfection / Q000379:methods*
chemical_list: C476399:Cd44 protein, mouse; C554870:Cd9 protein, mouse; D018398:Homeodomain Proteins; D018960:Hyaluronan Receptors; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D016271:Proto-Oncogene Proteins c-myc; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D060245:Tetraspanin 29; C495971:Hras protein, mouse; D016283:Proto-Oncogene Proteins p21(ras)