Increased level of exogenous Zinc induces cytotoxicity and up-regulates the expression of the ZnT-1 Zinc transporter gene in pancreatic cancer cells

Department of Surgery, College of Medicine, University of Illinois at Chicago, Chicago, IL 60612, USA.
The Journal of nutritional biochemistry (Impact Factor: 3.79). 04/2010; 22(1):79-88. DOI: 10.1016/j.jnutbio.2009.12.001
Source: PubMed


A balance between zinc uptake by ZIP (SLC39) and efflux of zinc from the cytoplasm into subcellular organelles and out of the cell by ZnT (SLC30) transporters is crucial for zinc homeostasis. It is not clear whether normal and cancerous pancreatic cells respond differently to increased extracellular zinc concentrations. Use of flow cytometry-based methods revealed that treatment with as little as 0.01 mM zinc induced significant cytotoxicity in two human ductal adenocarcinoma cell lines. In contrast, normal human pancreatic islet cells tolerated as high as 0.5 mM zinc. Insulinoma cell lines of mouse and rat origin also succumbed to high concentrations of zinc. Exposure to elevated zinc concentrations enhanced the numbers of carcinoma but not primary islet cells staining with the cell-permeable zinc-specific fluorescent dye, FluoZin-3, indicating increased zinc influx in transformed cells. Mitochondrial membrane depolarization, superoxide generation, decreased antioxidant thiols, intracellular acidosis and activation of intracellular caspases characterized zinc-induced carcinoma cell death. Only the antioxidant glutathione but not inhibitors of enzymes implicated in apoptosis or necrosis prevented zinc-induced cytotoxicity in insulinoma cells. Immunoblotting revealed that zinc treatment increased the ubiquitination of proteins in cancer cells. Importantly, zinc treatment up-regulated the expression of ZnT-1 gene in a rat insulinoma cell line and in two human ductal adenocarcinoma cell lines. These results indicate that the exposure of pancreatic cancer cells to elevated extracellular zinc concentrations can lead to cytotoxic cell death characterized by increased protein ubiquitination and up-regulation of the zinc transporter ZnT-1 gene expression.

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Available from: Sundararajan Jayaraman,
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    • "We initially sought to investigate MTF-1 regulated proteins by 2DE. However, we obtained no evidence on our 2DE gels for those proteins known to be regulated by MTF-1 and thought to be the key proteins in maintaining zinc homeostasis, such as MTs, ZnTs or ZIPs [15], [30], [46], [47]. To explore possible reasons for this, we measured the abundance of influx transporter ZIP-1, efflux transporter ZnT-1, zinc binding protein MT-1 and MTF-1 in A549 cells after treatment with 100 µM ZnSO4 in a time-dependent manner. "
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