Alterations of gene expression in endoderm differentiation of F9 teratocarcinoma cells.
During the endoderm differentiation of F9 mouse embryonal carcinoma cells, as induced by sodium butyrate (NaBu) or retinoic acid (RA), gene expressions of alkaline phosphatase (ALPase), pyruvate kinase (PKase) and 5' ribonucleotide phosphohydrolase (5'-Nase) were examined. The specific activity of ALPase was found to increase by 3.5-fold after 48 hr treatment with NaBu. In contrast, specific activity of PKase were decreased by 63%. Northern blot analysis revealed that the elevation of ALPase activity resulted from an increase in the level of liver/bone/kidney (L/B/K)-type ALPase mRNA and that the decrease of PKase activity was dependent on a reduction in the level of M(2)-PKase mRNA. Interestingly, when NaBu was removed from the culture medium, the levels of these mRNAs reverted to their original levels after 16 h. During these processes, the specific activity of 5'-Nase and the level of its mRNA remained unchanged. In contrast, when F9 cells were treated with RA, only the level of L/B/K-type ALPase mRNA increased. Lastly, we examined the issue of whether an increase in the level of ALPase mRNA is dependent on the transcriptional activation of the mouse L/B/K-type ALPase gene. Transient transfection assays using luciferase reporter constructs showed that the promoter activity increased as the result of treatment with RA but not with NaBu.
Shimada
N
N
Department of Biology, Osaka Kyoiku University, Osaka, Japan.
Yamada
K
K
Tanaka
T
T
Kawata
H
H
Mizutani
T
T
Miyamoto
K
K
Matsuzawa
T
T
D015720:5'-Nucleotidase / Q000235:genetics / Q000378:metabolism*; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D002454:Cell Differentiation / Q000235:genetics*; D004707:Endoderm / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology*; D015870:Gene Expression; D017930:Genes, Reporter / Q000235:genetics; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics; D011770:Pyruvate Kinase / Q000235:genetics / Q000378:metabolism*; D018243:Teratocarcinoma; D014407:Tumor Cells, Cultured
D002087:Butyrates; D011770:Pyruvate Kinase; D000469:Alkaline Phosphatase; D015720:5'-Nucleotidase