title: Gallium-induced cell death in lymphoma: role of transferrin receptor cycling, involvement of Bax and the mitochondria, and effects of proteasome inhibition.
abstract: Gallium nitrate is a metallodrug with clinical efficacy in non-Hodgkin's lymphoma. Its mechanisms of antineoplastic action are not fully understood. In the present study, we investigated the roles of transferrin receptor (TfR) targeting and apoptotic pathways in gallium-induced cell death. Although DoHH2 lymphoma cells displayed a 3-fold lower number of TfRs than CCRF-CEM lymphoma cells, they were 3- to 4-fold more sensitive to gallium nitrate. Despite a lower TfR expression, DoHH2 cells had greater TfR cycling and iron and gallium uptake than CCRF-CEM cells. In other lymphoma cell lines, TfR levels per se did not correlate with gallium sensitivity. Cells incubated with gallium nitrate showed morphologic changes of apoptosis, which were decreased by the caspase inhibitor Z-VAD-FMK and by a Bax-inhibitory peptide. Cells exposed to gallium nitrate released cytochrome c from mitochondria and displayed a dose-dependent increase in caspase-3 activity. An increase in active Bax levels without accompanying changes in Bcl-2 or Bcl-X(L) was seen in cells incubated with gallium nitrate. The endogenous expression of antiapoptotic Bcl-2 was greater in DoHH2 cells than in CCRF-CEM cells, suggesting that endogenous Bcl-2 levels do not correlate with cell sensitivity to gallium nitrate. Gallium-induced apoptosis was enhanced by the proteasome inhibitor bortezomib. Our results suggest that TfR function rather than TfR number is important in gallium targeting to cells and that apoptosis is triggered by gallium through the mitochondrial pathway by activating proapoptotic Bax. Our studies also suggest that the antineoplastic activity of combination gallium nitrate and bortezomib warrants further investigation.
authors.affiliation: Division of Neoplastic Diseases, Department of Medicine, Medical College of Wisconsin, 9200 West Wisconsin Avenue, Milwaukee, WI 53226. chitambr@mcw.edu.
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Chitambar
authors.forename: Janine P
authors.initials: JP
authors.lastname: Wereley
authors.forename: Shigemi
authors.initials: S
authors.lastname: Matsuyama
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology / Q000633:toxicity*; D053148:Caspase 3 / Q000378:metabolism; D061945:Caspase Inhibitors; D016923:Cell Death; D045304:Cytochromes c / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005708:Gallium / Q000494:pharmacology / Q000633:toxicity*; D006801:Humans; D007501:Iron / Q000378:metabolism; D019169:Jurkat Cells; D008228:Lymphoma, Non-Hodgkin / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D008928:Mitochondria / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000494:pharmacology; D011990:Receptors, Transferrin / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured; D051028:bcl-2-Associated X Protein / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D061945:Caspase Inhibitors; D011990:Receptors, Transferrin; D051028:bcl-2-Associated X Protein; D045304:Cytochromes c; D005708:Gallium; D007501:Iron; D053148:Caspase 3; D046988:Proteasome Endopeptidase Complex; C027235:gallium nitrate