Nuclear and cytoplasmic expressions of the carbohydrate-binding protein CBP70 in tumoral or healthy cells of the macrophagic lineage.
The expression of the carbohydrate-binding protein CBP70 was analyzed in undifferentiated HL60 cells, HL60 cells differentiated into monocytes/macrophages or granulocytes, healthy monocytes and in vitro monocyte-derived macrophages (MDM) using an anti-CBP70 serum. This study was performed by immunoblotting analysis of nuclear and cytoplasmic extracts before and after N-acetylglucosamine affinity chromatography and by indirect immunofluorescence microscopy. The results of this study show, for the first time, that CBP70 is expressed in the nucleus and the cytoplasm of healthy or leukemic cells of the macrophagic lineage. However, striking differences were observed depending upon the leukemic or normal state of cells and cell differentiation. Indeed, the level of expression and the intracellular distribution of CBP70 were found to be different in undifferentiated HL60 cells and monocytes/macrophages differentiated from these cells. Major differences were also observed according to whether macrophages differentiated from leukemic HL60 cells or healthy monocytes. Thus, the total cellular expression of CBP70 was markedly lower in MDM than in HL60-derived macrophages and the intracellular distribution of the protein was different. Nevertheless, in both cases, the total cellular expression of CBP70 was enhanced during cell differentiation. Another important result is the finding that CBP70 behaviour was totally different when HL60 cells were induced to differentiate into macrophages or granulocytes. These data could therefore suggest that CBP70 is involved in phagocytic cell differentiation. Moreover, we show that an additional 60 kDa polypeptide (p60), recognized by the anti-CBP70 serum, is expressed in HL60 cells differentiated into macrophages or granulocytes as well as in healthy monocytes or MDM but not expressed in undifferentiated HL60 cells. Although CBP70 and p60 appeared to be closely related polypeptides, their relationship remains to be precised. These findings are discussed with regard to data available on galectin-3.
Hadj Sahraoui
Y
Y
INSERM U180, Centre Universitaire des Saints-Pères, Université René-Descartes, Paris, France.
Sève
A P
AP
Doyennette-Moyne
M A
MA
Saffar
L
L
Felin
M
M
Aubery
M
M
Gattegno
L
L
Hubert
J
J
D002454:Cell Differentiation; D002467:Cell Nucleus / Q000737:chemistry; D002470:Cell Survival; D003593:Cytoplasm / Q000737:chemistry; D005434:Flow Cytometry; D018922:HL-60 Cells; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D008264:Macrophages / Q000737:chemistry* / Q000166:cytology; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D014407:Tumor Cells, Cultured
D037102:Lectins; C104787:carbohydrate-binding protein 70, human