Resistance to the Antineoplastic Agent Gallium Nitrate Results in Marked Alterations in Intracellular Iron and Gallium Trafficking: Identification of Novel Intermediates

Abstract

Gallium (Ga) shows significant antitumor activity by markedly interfering with iron (Fe) metabolism, and (67)Ga is used as a radio-imaging agent for cancer detection. Therefore, the mechanisms involved in (67)Ga uptake, metabolism, and resistance are critical to understand. The development of tumor lines that are gallium-resistant suggests (67)Ga uptake may be different in these cells, providing an opportunity for understanding intracellular (67)Ga and (59)Fe transport and gallium resistance. In this study, gallium-resistant cells were used to assess (67)Ga and (59)Fe uptake using native polyacrylamide gel electrophoresis autoradiography. In contrast to the common view that (67)Ga and (59)Fe use the same uptake pathways, we show that the trafficking of these two metal ions is different in cells either resistant (R) or sensitive (S) to gallium. Indeed, in contrast to (59)Fe, little (67)Ga is incorporated into ferritin, with most present as a labile (67)Ga pool. We also report unique changes in (67)Ga and (59)Fe trafficking between R and S cells. In particular, in R cells, there was a distinct transferrin-transferrin receptor 1-hemochromatosis protein (HFE) complex (band B) not observed in S cells. Furthermore, because HFE regulates iron and gallium uptake, the two Tf-TfR1-HFE complexes in R cells may be involved in reduced (67)Ga and (59)Fe uptake compared with S cells. In S cells, a novel iron-binding intermediate (band D) was identified that was not present in R cells and may be a "sensitivity factor" to gallium. In contrast to the general view that (67)Ga and (59)Fe use the same or similar uptake pathways, we show that their distribution and trafficking is markedly different in R and S cells.