Specific effect of 5-fluorouracil on alpha-fetoprotein gene expression during the in vitro mouse embryonic stem cell differentiation.
Embryonic stem (ES) cells are considered an important alternative to develop in vitro screening methods for embryotoxicity. Mouse ES cells can be cultured as cell suspension aggregates termed "embryoid bodies" (EBs) in which cells start to differentiate. We have studied the expression of several genes in the presence of a wide range of concentrations of 5-fluorouracil (5-FU). This well-established embryotoxic compound completely inhibited cell viability at 200 nmol/L in monolayer cultures. At lower concentrations, 5-FU led to decrease in the expression of the alpha-fetoprotein gene, a marker of the visceral endoderm, in the EBs. However, the expression of several mesodermal gene markers was not significantly affected at these concentrations. These results suggest a high sensitivity of the visceral endoderm differentiation to 5-FU. Therefore, the quantification of the alpha-fetoprotein gene after exposure to potential embryotoxicants should be considered an additional end point in future embryotoxicity assays in vitro with ES cells.
Pamies
David
D
Instituto de Bioingenieria, Universidad Miguel Hernández de Elche, Elche, Spain.
Vicente-Salar
Néstor
N
Sogorb
Miguel A
MA
Roche
Enrique
E
Reig
Juan A
JA
D000818:Animals; D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004707:Endoderm / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D023381:Endpoint Determination; D005472:Fluorouracil / Q000633:toxicity*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism; D013723:Teratogens / Q000633:toxicity*; D013997:Time Factors; D018675:Toxicity Tests / Q000379:methods; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000235:genetics / Q000378:metabolism*; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism
C424167:Amn protein, mouse; D000943:Antigens, Differentiation; C495267:CTNNB1 protein, mouse; D008565:Membrane Proteins; C551613:Myh7 protein, mouse; D013723:Teratogens; D000509:alpha-Fetoproteins; D051176:beta Catenin; D040301:Vascular Endothelial Growth Factor Receptor-2; D018995:Myosin Heavy Chains; D005472:Fluorouracil