Differential regulation of vitamin D receptors in clonal populations of a chronic myelogenous leukemia cell line.
RWLeu4 is a chronic myelogenous leukemia cell line that is sensitive to the antiproliferative and differentiation-inducing actions of 1alpha,25(OH)2-vitamin D3 (VD3). The JMRD3 cell line is a VD3-resistant variant of RWLeu4 that was selected by continuous passage of RWLeu4 in the presence of VD3. The isolation of a spontaneous VD3-resistant variant suggests that phenotypically different cells exist within the RWLeu4 cell population. Therefore, single-cell clones of RWLeu4 cells were isolated and characterized. Four clonal cell populations that fall into three groups differing in response to the antiproliferative and differentiation-inducing actions of VD3 were examined. Surprisingly, the extent of response of the clones to VD3 does not show a correlation with the basal level of the vitamin D receptor (VDR). RWLeu4-3 and RWLeu4-4 are the clones most sensitive to the antiproliferative actions of VD3 (ED50 approximately equal to 1 nM); however, RWLeu4-3 expresses basal levels of VDRs similar to those found in the parental cells and the RWLeu4-2 clone, while in RWLeu4-4, VD3 binding and VDR protein are below the limits of detection. Furthermore, RWLeu4-10 expresses the highest basal level of VDR protein but is relatively resistant to the antiproliferative actions of VD3 (ED50 > or = 30 nM). Like JMRD3, RWLeu4-10 is still capable of differentiating in response to VD3, as judged by the induction of biochemical processes and cell-surface antigen expression. Although VD3 treatment increases VDR protein levels and DNA-binding activity in all clones, altered DNA-protein complexes are detected in RWLeu4-4. Our results suggest that sensitivity to the antiproliferative and differentiation-inducing actions of VD3 is not dependent solely upon the level of VDR expressed, but may also require posttranslational modification of the VDR or complex interactions with other nuclear transcription factors.
Iwata
K
K
Roger Williams Medical Center, Experimental Pathology Section, Department of Pathology and Laboratory, Brown University School of Medicine, Providence, RI 02908, USA. stephen_lasky@brown.edu
Kouttab
N
N
Ogata
H
H
Morgan
J W
JW
Maizel
A L
AL
Lasky
S R
SR
D001483:Base Sequence; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002454:Cell Differentiation; D002762:Cholecalciferol / Q000494:pharmacology*; D002999:Clone Cells; D004247:DNA / Q000378:metabolism; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D016177:Macrophage-1 Antigen / Q000378:metabolism; D008969:Molecular Sequence Data; D009580:Nitroblue Tetrazolium / Q000378:metabolism; D009838:Oligodeoxyribonucleotides; D018167:Receptors, Calcitriol / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D018950:Lipopolysaccharide Receptors; D016177:Macrophage-1 Antigen; D009838:Oligodeoxyribonucleotides; D018167:Receptors, Calcitriol; D002762:Cholecalciferol; D009580:Nitroblue Tetrazolium; D004247:DNA; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase