Gene expression in hepatocyte-like lines established by targeted carcinogenesis in transgenic mice.
New hepatocyte-like cell lines (mhAT) were derived from the liver of a transgenic mouse expressing SV40 early genes under the direction of the liver-specific antithrombin III gene promoter (ATIII-TSV40). Their differentiated phenotypes were improved and stabilized by the use of liver-specific growth media (arginine-free, glucose-free, or low-fructose/glucose-free medium). The best differentiated lines display a very high level of albumin, transferrin, and L-type pyruvate kinase (L-PK) gene expression that is comparable to that observed in the mouse liver. Abundance of the aldolase B and phosphoenolpyruvate carboxykinase (PEPCK) transcripts varied from 5 to 35% of the in vivo concentrations while abundance of the alpha-fetoprotein and phenylalanine hydroxylase transcripts remained very low. Hormonal (cAMP and insulin) and nutritional (glucose) gene controls of PEPCK and L-PK were, at least partially, conserved. mhAT cells are readily transfectable by the calcium phosphate coprecipitation technique and exhibit a liver-specific pattern of expression of exogenous genes. Thus, mhAT cells seem suitable for the analysis of the regulatory regions involved in the tissue-specific transcription of genes. This work demonstrates, therefore, the great efficiency of targeted carcinogenesis in transgenic mice to create new differentiated cell lines. The availability of various lines of liver-specific cells with different phenotypes will constitute useful tools to establish correlations between expression of trans-acting factors and control of the phenotype.
Antoine
B
B
Institut Cochin de Génétique Moléculaire, laboratoire de recherche en génétique et pathologie moléculaires (INSERM U129), Paris, France.
Levrat
F
F
Vallet
V
V
Berbar
T
T
Cartier
N
N
Dubois
N
N
Briand
P
P
Kahn
A
A
D000418:Albumins / Q000032:analysis; D000818:Animals; D000957:Antigens, Viral, Tumor / Q000235:genetics; D002460:Cell Line / Q000378:metabolism*; D005786:Gene Expression Regulation; D008099:Liver / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D010641:Phenotype; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000235:genetics; D011401:Promoter Regions, Genetic; D011770:Pyruvate Kinase / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D015534:Trans-Activators; D014162:Transfection; D014168:Transferrin / Q000032:analysis; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins / Q000032:analysis
D000418:Albumins; D000957:Antigens, Viral, Tumor; D012333:RNA, Messenger; D015534:Trans-Activators; D014168:Transferrin; D000509:alpha-Fetoproteins; D011770:Pyruvate Kinase; D010729:Phosphoenolpyruvate Carboxykinase (GTP)