Auranofin an Anti-Rheumatic Gold Compound Modulates Apoptosis by Elevating the Intracellular Calcium Concentration (Ca2+i) in MCF-7 Breast Cancer Cells

摘要

Auranofin, a transition metal complex is used for the treatment of rheumatoid arthritis but is also an effective anti-cancer drug. We investigate the effects of Auranofin in inducing cell death by apoptosis and whether these changes are correlated to changes of intracellular calcium concentration ([Ca²⁺]i) in breast cancer cells (MCF-7). Cytotoxicity of Auranofin was evaluated using MTS assay and the Trypan blue dye exclusion method. With fluorescent dyes SR-FLICA and 7-AAD apoptotic death and necrotic death were differentiated by Flow cytometry. A concentration dependent decrease in the viability occurred and cells were shifted to the apoptotic phase. Intracellular calcium ([Ca²⁺]i) was recorded using florescence microscopy and a calcium sensitive dye (Fluo-4 AM) with a strong negative correlation (r = −0.713) to viability. Pharmacological modulators 2-APB (50 μM), Nimodipine (10 μM), Caffeine (10 mM), SKF 96365(20 μM) were used to modify calcium entry and release. Auranofin induced a sustained increase of [Ca²⁺]i in a concentration and time dependent manner. The use of different blockers of calcium channels did not reveal the source for the rise of [Ca²⁺]i. Overall, elevation of [Ca²⁺]i by Auranofin might be crucial for triggering Ca²⁺-dependent apoptotic pathways. Therefore, in anti-cancer therapy, modulating [Ca²⁺]i should be considered as a crucial factor for the induction of cell death in cancer cells.