The apolipoprotein B gene is constitutively expressed in HepG2 cells: regulation of secretion by oleic acid, albumin, and insulin, and measurement of the mRNA half-life.
The objective of this study was to establish conditions whereby apoB secreted from HepG2 cells could be regulated over a wide range, and to determine whether changes of output were correlated with the level of apoB mRNA. The presence of oleate (complexed to 3% albumin at a molar ratio of 1.7:1) resulted in a 3.5-fold stimulation of apoB secretion that was apparent after only 3 h. Insulin halved the rate of apoB output and the inhibition was detectable within the physiological insulin range, but was not apparent until 12-16 h. Albumin in the culture medium had a dose-dependent inhibitory effect on apoB production. Overall, apoB secretion from HepG2 cells was modulated over a 7-fold range. However, when apoB mRNA was assayed by slot-blot hybridization, no change was detectable under any of the conditions that modulated apoB output. Quantitative solution hybridization was used to confirm that oleate did not affect the level of apoB mRNA. Kinetic analysis of the decay of [3H]uridine-labeled apoB mRNA showed that the half-life of apoB mRNA was 16 h. We conclude from these studies that the apoB gene is constitutively expressed in HepG2 cells and that the mechanism of acute regulation of apoB production by these cells must involve co- or post-translational processes.
Pullinger
C R
CR
Division of Molecular Medicine, MRC Clinical Research Centre, Harrow, United Kingdom.
North
J D
JD
Teng
B B
BB
Rifici
V A
VA
Ronhild de Brito
A E
AE
Scott
J
J
D000418:Albumins / Q000494:pharmacology*; D001055:Apolipoproteins B / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D015152:Blotting, Northern; D015342:DNA Probes; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation; D006207:Half-Life; D006801:Humans; D007328:Insulin / Q000494:pharmacology*; D019301:Oleic Acid; D009829:Oleic Acids / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000418:Albumins; D001055:Apolipoproteins B; D015342:DNA Probes; D007328:Insulin; D009829:Oleic Acids; D012333:RNA, Messenger; D019301:Oleic Acid