A change in chromosome number, known as aneuploidy, is a common characteristic of cancer. Aneuploidy disrupts gene expression in human cancer cells and immortalized human epithelial cells, but not in normal human cells. However, the relationship between aneuploidy and cancer remains unclear. To study the effects of aneuploidy in normal human cells, we generated artificial cells of human primary fibroblast having three chromosome 8 (trisomy 8 cells) by using microcell-mediated chromosome transfer technique. In addition to decreased proliferation, the trisomy 8 cells lost contact inhibition and reproliferated after exhibiting senescence-like characteristics that are typical of transformed cells. Furthermore, the trisomy 8 cells exhibited chromosome instability, and the overall gene expression profile based on microarray analyses was significantly different from that of diploid human primary fibroblasts. Our data suggest that aneuploidy, even a single chromosome gain, can be introduced into normal human cells and causes, in some cases, a partial cancer phenotype due to a disruption in overall gene expression.

Superoxide dismutases (SODs) are antioxidant proteins that convert superoxide to hydrogen peroxide. In vertebrate cells, SOD1 is mainly present in the cytoplasm, with small levels also found in the nucleus and mitochondrial intermembrane space, and SOD2 is present in the mitochondrial matrix. Previously, the authors conditionally disrupted the SOD1 or SOD2 gene in DT40 cells and found that depletion of SOD1 caused lethality, while depletion of SOD2 led to growth retardation. The observations from previous work showed that the lethality observed in SOD1-depleted cells was completely rescued by ascorbic acid. Ascorbic acid is a water-soluble antioxidant present in biological fluids; however, the exact target for its antioxidant effects is not known. In this study, the authors demonstrated that ascorbic acid offset growth defects observed in SOD2-depleted cells and also lowered mitochondrial superoxide to physiological levels in both SOD1- or SOD2-depleted cells. Moreover, depletion of SOD1 or SOD2 resulted in the accumulation of intracellular oxidative stress, and this increased oxidative stress was reduced by ascorbic acid. Taken together, this study suggests that ascorbic acid can be applied as a nontoxic antioxidant that mimics the functions of cytoplasmic and mitochondrial SODs.

The effects of high glucose concentrations on the selective permeability of a layer of cultured bovine vascular endothelial cells (ECs) were investigated. ECs were derived from the intima of the thoracic aorta and permeability to nonglycated albumin, glycated albumin, and fluorescein dextrans (FDs) of molecular mass to albumin was measured. ECs were cultured on a filter coated with type I collagen and preincubated in the presence of various glucose concentrations for 96 h. Human serum albumin was glycated by incubation with glucose in vitro. Nonglycated and glycated albumin were separated by affinity column chromatography. The permeation rates of nonglycated and glycated albumin as well as those of neutral and anionic FD through the EC layer were increased by preincubation of cells with high glucose concentrations (22.2 and 44.4 mmol/l). The permeation rate of glycated albumin was significantly less than that of nonglycated albumin at all glucose concentrations tested, whereas the permeation rate of anionic FD was significantly lower than that of neutral FD only at a physiological glucose concentration (5.6 mmol/l). The aldose reductase inhibitor ponalrestat partially inhibited the high glucose-induced increase in trans-endothelial permeation of albumin (both nonglycated and glycated), but had no effect on the increased permeation of FD. These results indicate that high glucose concentrations enhance trans-endothelial permeability to albumin and FD and may disturb the barrier function of vascular ECs. Furthermore, metabolism of glucose via the polyol pathway may contribute to abnormalities in trans-endothelial permeability.

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