Effects of actinomycin D and cycloheximide on transcript levels of IGF-I, actin, and albumin in hepatocyte primary cultures treated with growth hormone and insulin.
The stability of several RNA transcripts in cultured hepatocytes is known to increase when serum is omitted from the culture medium. In order to investigate possible mechanisms for this phenomenon, we examined the effects of actinomycin D and cycloheximide on the levels of actin, albumin, and insulin-like growth factor I transcripts in primary cultures incubated in serum-free medium. The levels of IGF-I and albumin transcripts per culture increased for the first 4 hours following addition of actinomycin D and then declined. The levels of actin transcripts and total RNA per culture declined immediately following actinomycin D addition in a manner consistent with exponential decay. IGF-I and albumin transcript levels were relatively unaffected by cycloheximide, while actin transcript levels increased 7-fold over 7 hours. The half-lives of actin transcripts and total RNA were calculated to be 4.6 to 7.7 hours and 11 to 19 hours, respectively, with no statistically significant correlation with hormone treatment. The data suggest that the stability of albumin and IGF-I transcripts, but not actin transcripts, is controlled in part by an actinomycin D-sensitive process.
Johnson
T R
TR
Department of Anatomy, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106.
Trojan
J
J
Rudin
S D
SD
Blossey
B K
BK
Ilan
J
J
Ilan
J
J
D000199:Actins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D000418:Albumins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D003470:Culture Media; D003513:Cycloheximide / Q000494:pharmacology*; D003609:Dactinomycin / Q000494:pharmacology*; D005260:Female; D013006:Growth Hormone / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000378:metabolism; D012313:RNA / Q000032:analysis* / Q000235:genetics; D051381:Rats; D011919:Rats, Inbred Strains; D014158:Transcription, Genetic
D000199:Actins; D000418:Albumins; D003470:Culture Media; D007328:Insulin; D003609:Dactinomycin; D012313:RNA; D007334:Insulin-Like Growth Factor I; D013006:Growth Hormone; D003513:Cycloheximide