Transferrin iron interactions with cultured hepatocellular carcinoma cells (PLC/PRF/5).
Hepatocellular carcinoma cells of the PLC/PRF/5 cell line had 1.9 x 10(5) transferrin receptors per tumor cell with a Kd of 1.5 x 10(-8) M. At high concentrations of transferrin the binding was not saturable. Transferrin internalization by hepatoma cells was shown by time and temperature-dependent binding studies and by pronase experiments. Transferrin recycling was confirmed by the demonstration of a progressive increase in the cellular molar ratios of iron to transferrin and by chase experiments. Ammonium chloride interfered with iron unloading. The vinca alkaloid vincristine inhibited iron and transferrin uptake. The hepatocarcinoma cells appeared to lack asialoglycoprotein receptors and therefore internalized partially desialated transferrin by the regular route. Iron uptake from transferrin was markedly inhibited by the hydrophobic ferrous chelator 2,2' bipyridine but was relatively unaffected by the hydrophilic ferric chelator desferroxamine. The implication that ferrous iron was involved in postendocytic transvesicular membrane iron transport was supported by a study in which hepatoma cells were shown to take up large amounts of ferrous iron suspended in 270 mM sucrose at pH 5.5. The interaction at this pH between surface labeled hepatoma cell extracts and ferrous iron on a Sephacryl S-300 column suggested that the postendocytic transvesicular transport of iron through the membrane was in part protein mediated. The endocytosed iron in hepatoma cells was found in association with ferritin (33%), transferrin (31%) and a low molecular weight fraction (21%).
Baynes
R
R
Department of Medicine, University of the Witwatersrand Medical School, Johannesburg, South Africa.
Friedman
B
B
McNamara
L
L
Bothwell
T
T
Bukofzer
G
G
Macfarlane
B
B
D015082:2,2'-Dipyridyl / Q000494:pharmacology; D000818:Animals; D003676:Deferoxamine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis; D006801:Humans; D007501:Iron / Q000493:pharmacokinetics*; D008114:Liver Neoplasms, Experimental / Q000032:analysis / Q000378:metabolism; D013997:Time Factors; D014168:Transferrin / Q000378:metabolism / Q000493:pharmacokinetics*; D014407:Tumor Cells, Cultured / Q000032:analysis / Q000378:metabolism*
D014168:Transferrin; C028283:diferric transferrin; D015082:2,2'-Dipyridyl; D007501:Iron; D003676:Deferoxamine