Characterization of cadmium uptake in human intestinal crypt cells HIEC in relation to inorganic metal speciation.
Cadmium (Cd) uptake was studied under inorganic exposure conditions in normal human intestinal crypt cells HIEC. The uptake time course of 0.3 microM Cd in a serum-free chloride medium was analyzed according to a first order equation with rapid initial (U0) and maximal (Umax) accumulation values of 14.1+/-1.4pmol/mgprotein and 41.4+/-2.0 pmol/mgprotein, respectively. The presence of a 300-fold excess of unlabeled Cd dramatically decreased tracer uptake, showing the involvement of specific mechanism(s) of transport. Our speciation studies revealed the preferential uptake of the free ion Cd2+, but also suggested that CdCln(2-n) species may contribute to Cd accumulation. Specific mechanisms of transport of very high and similar affinity (Km approximately 5 microM) have been characterized under both chloride and nitrate exposure conditions, but a two-fold higher capacity (Vmax) was estimated in the nitrate medium used to increase [Cd2+] over chlorocomplex formation. A clear inhibition of 109Cd uptake was observed at external acidic pH under both exposure media. An La-inhibitible 46% increase in 109Cd uptake was obtained in nominally Ca-free nitrate medium, whereas Zn provided additional inhibition. These results show different kinetic parameters for Cd uptake as a function of inorganic metal speciation. Cd2+ uptake would not involve the H+-coupled symport NRAMP2 but would be related instead to the Ca and/or Zn pathways. Because proliferative crypt cells play a critical role in the renewal process of the entire intestinal epithelium, studies on the impact of Cd on HIEC cell functions clearly deserve further investigation.
Bergeron
Pierre-Michel
PM
Département des Sciences Biologiques, Centre TOXEN, Université du Québec à Montréal, C.P. 8888, Succ. Centre-ville, Montréal, Que., Canada H3C 3P8.
Jumarie
Catherine
C
D000465:Algorithms; D018938:Caco-2 Cells; D002104:Cadmium / Q000378:metabolism*; D002106:Cadmium Radioisotopes; D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000096:biosynthesis; D019073:HT29 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007413:Intestinal Mucosa / Q000378:metabolism*; D007422:Intestines / Q000166:cytology; D033862:Iron-Binding Proteins / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008670:Metals / Q000737:chemistry / Q000494:pharmacology*; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D012044:Regression Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000099236:Solute Carrier Family 11, Member 2
D002104:Cadmium; D002106:Cadmium Radioisotopes; D027682:Cation Transport Proteins; D018076:DNA, Complementary; D033862:Iron-Binding Proteins; D008670:Metals; D012313:RNA; D000099236:Solute Carrier Family 11, Member 2