Cadmium Uptake and Transepithelial Transport in Control and Long-Term Exposed Caco-2 Cells: the Role of Metallothionein

Exposure of humans to cadmium, a common environmental pollutant, is mainly through food intake. However, the mechanisms of intestinal absorption have not been clearly elucidated for this toxic metal ion. In order to investigate the effects of long-term exposure to this metal and the role of metallothioneins in cadmium absorption, we used human-derived Caco-2 cells cultured on porous membrane filters. We first validated this model by quantifying metal uptake and transepithelial transport on control cells and cells adapted to grow for 2 to 5 weeks in the presence of low doses of cadmium in the culture medium. The nontoxic doses of cadmium (0.1, 1.0, and 5 μM), in which Caco-2 cells could be cultured for many passages without deleterious effects, were determined by evaluating transepithelial resistance of the cells and lactate dehydrogenase leakage. After 24 h of 1 μM Cd exposure, intracellular cadmium levels were 3- and 6-fold higher for cells exposed for extended periods to 1 and 5 μM cadmium, respectively, compared to control cells. In control and long-term exposed cells, this accumulation was inhibited by zinc, copper, and pCMBS, but not by verapamil or ouabain. Intracellular metallothionein content was increased 1.5-, 5-, and 12-fold for the cells grown in the presence of 0.1, 1.0, and 5 μM cadmium, respectively, in the culture medium. The amount of metallothionein synthesized and released by the cells was highly correlated with cadmium accumulation and transport. Our results suggest that Caco-2 cell monolayers are a good predictive model for the study of cadmium intestinal absorption following exposure to repeated low doses of cadmium, and confirm the essential role of metallothionein in the regulation of cadmium intestinal absorption.