Production and secretion of retinol-binding protein by a human hepatoma cell line, HepG2.
Retinol-binding protein (RBP) that is synthesized and secreted by the human hepatoma cell HepG2 has been measured using a sensitive radioimmunoassay in which RBP in media and hepatoma cell sonicates reacts identically to human serum RBP. RBP was synthesized and secreted when cells were grown in retinol-depleted as well as retinol-containing media. However, immunoreactive transthyretin (prealbumin) could not be detected in concentrated HepG2 medium. RBP secretion and accumulation per mg of cell protein could be modulated by the concentration of fetal calf serum in the growth medium: secreted RBP equaled 782 +/- 123 ng/mg of cell protein per 8 hr after preincubation with 10% fetal calf serum versus 555 +/- 86 ng/mg per 8 hr in the absence of serum, whereas RBP in cell sonicates decreased only slightly. When HepG2 cells were cultured for two or more passages in medium containing fetal calf serum depleted of retinol by ultraviolet irradiation, the amounts of RBP in the cells and released to the medium were both significantly increased. When vitamin A (90% as retinyl esters) in the form of chylomicron remnants was presented to cells, there was a significant, dose-dependent redistribution of RBP from cells to medium, both in cells grown in normal fetal calf serum and in retinol-depleted serum. These data indicate that the secretion of RBP by HepG2 can occur constitutively in the absence of retinol, but that secretion can be enhanced and regulated by retinol delivered by the chylomicron remnant.
Marinari
L
L
Department of Physiology and Biochemistry, Medical College of Pennsylvania, Philadelphia 19129.
Lenich
C M
CM
Ross
A C
AC
D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D002460:Cell Line; D002914:Chylomicrons / Q000494:pharmacology; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism*; D011863:Radioimmunoassay; D012177:Retinol-Binding Proteins / Q000096:biosynthesis* / Q000378:metabolism; D014801:Vitamin A / Q000494:pharmacology
D002914:Chylomicrons; D012177:Retinol-Binding Proteins; D014801:Vitamin A