Characterization of nuclear retinoic acid binding activity in sensitive leukemic cell lines: cell specific uptake of ATRA and RAR alpha protein modulation.
The diverse effects of all-trans retinoic acid (ATRA) on growth, differentiation and homeostasis of vertebrate organisms are mediated by three distinct isoforms of retinoic acid receptors (RARs). Although it is not known to what extent each RAR contributes to the different effects of ATRA, several studies have demonstrated that ATRA induced granulocytic differentiation in human myeloid leukemic cell lines is mediated by RAR alpha. In this study, we investigated ATRA binding affinity of the endogenous nuclear receptors of HL-60 and NB4 leukemic cells. Scatchard plot analysis yielded an apparent dissociation constant of 5 +/- 0.3 nM and 1400 +/- 80 receptor sites per cell in HL-60 cells, whereas the NB4 promyelocytic leukemic cell line showed a lower affinity (8.5 +/- 0.5 nM and 900 +/- 30 receptor sites per cell). Modulation of RAR alpha protein (5 fold excess) was found in NB4 cells after 24 hours ATRA exposure, whereas HL-60 cells required a 72-hour culture period to weakly increase the RAR alpha protein level. These data were closely related to the ATRA intracellular concentration and kinetics of terminal differentiation of the cells.
Agadir
A
A
Laboratoire de Biologie Cellulaire Hématopoïétique, Hôpital Saint-Louis, Paris, France.
Cornic
M
M
Jérôme
M
M
Menot
M L
ML
Cambier
N
N
Gaub
M P
MP
Gourmel
B
B
Lefebvre
P
P
Degos
L
L
Chomienne
C
C
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D001692:Biological Transport; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D006801:Humans; D007700:Kinetics; D015473:Leukemia, Promyelocytic, Acute; D008970:Molecular Weight; D018168:Receptors, Retinoic Acid / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D000072482:Retinoic Acid Receptor alpha; D014162:Transfection; D014212:Tretinoin / Q000378:metabolism* / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000911:Antibodies, Monoclonal; C000607071:RARA protein, human; D018168:Receptors, Retinoic Acid; D011994:Recombinant Proteins; D000072482:Retinoic Acid Receptor alpha; D014212:Tretinoin