A Proteomic View of Cellular Responses to Anticancer Quinoline-Copper Complexes

Abstract

Metal-containing drugs have long been used in anticancer therapies. The mechansims of action of platinum-based drugs are now well-understood, which cannot be said of drugs containing other metals, such as gold or copper. To gain further insights into such mechanisms, we used a classical proteomic approach based on two-dimensional elelctrophoresis to investigate the mechanisms of action of a hydroxyquinoline-copper complex, which shows promising anticancer activities, using the leukemic cell line RAW264.7 as the biological target. Pathway analysis of the modulated proteins highlighted changes in the ubiquitin/proteasome pathway, the mitochondrion, the cell adhesion-cytoskeleton pathway, and carbon metabolism or oxido-reduction. In line with these prteomic-derived hypotheses, targeted validation experiments showed that the hydroxyquinoline-copper complex induces a massive reduction in free glutathione and a strong alteration in the actin cytoskeleton, suggesting a multi-target action of the hydroxyquinoline-copper complex on cancer cells.

The longer the worse: a combined proteomic and targeted study of the long-termversusshort-term effects of silver nanoparticles on macrophages

“…To alleviate this problem, we used a spectrophotometric assay utilizing the cellular glutathione S-transferases and the chlorodinitrobenzene substrate. 38,39 After their exposure to silver in 6 well plates, cells were finally treated for 30 minutes with 25 μM chlorodinitrobenzene in the culture medium, resulting in the conjugation of glutathione to the substrate and consumption of the free reduced glutathione. 40,41 The culture medium was removed and the cell layer rinsed twice in Hepes NaOH buffer (10 mM pH 7.5) containing 2 mM magnesium acetate and 0.25 M sucrose.…”

Influences of Nanoparticles Characteristics on the Cellular Responses: The Example of Iron Oxide and Macrophages

“…Visualization of F-Actin cytoskeleton was assayed by phalloidin staining according to previously published protocols [38][39][40] 2.8. Glutathione Assays Intracellular glutathione levels were assessed using the monochlorobimane technique, with some modifications [33].…”

“…The mitochondrial transmembrane potential was assessed using the rhodamine 123 uptake assay, as previously described [40].…”

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