Characterization of the Na+/H+ exchanger in human choriocarcinoma (BeWo) cells.
We examined the expression and activity of the Na+/H+ exchanger in the human choriocarcinoma BeWo cell line. When treated with methotrexate, these cells differentiated from cytotrophoblast-like cells to enlarged multinucleate syncytiotrophoblast-like cells. There was no change in the apparent Km for Na+ between undifferentiated and differentiated cells. However, differentiated cells could transport more than five times the proton flux of undifferentiated cells. There was no difference in the Hill coefficient between undifferentiated and differentiated cells. However, the maximal flux (Jmax) for undifferentiated cells was higher than that for differentiated cells. Inhibition of Na+/H+ exchange activity by an amiloride analog and Hoe694 revealed a sensitive and a resistant component in both differentiated and undifferentiated cells. Northern blot analysis and immunocytochemistry suggested that the sensitive component was due to the NHE1 isoform of the protein while the resistant component was due to the NHE3 isoform. The NHE1 isoform was localized to the brush border membrane of BeWo cells and Western blot analysis showed that the NHE1 protein was more abundant in brush border membranes from differentiated BeWo cells compared to undifferentiated cells. The results show that BeWo cells contain the NHE1 and NHE3 isoforms of the Na+/H+ exchanger and that the NHE1 isoform is primarily localized to the brush border membrane.
Silva
N L
NL
Department of Pediatrics, Faculty of Medicine, University of Alberta, Edmonton, Canada.
Wang
H
H
Harris
C V
CV
Singh
D
D
Fliegel
L
L
D015152:Blotting, Northern; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002822:Choriocarcinoma / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007150:Immunohistochemistry; D007536:Isomerism; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D010641:Phenotype; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D017923:Sodium-Hydrogen Exchangers; D012313:RNA