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Normotonic cell shrinkage induces apoptosis under extracellular low Cl conditions in human lymphoid and epithelial cells

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Abstract Aim: 

Sustained cell shrinkage is associated with apoptosis. Apoptotic volume decrease, which is known to be induced by release of osmolytes including Cl ions, may be an essential event for apoptosis induction. Provided any anion channels and/or anion transporters are basally functioning, there is a possibility that imposition of a driving force for Cl efflux per se results in sustained cell shrinkage and thereby induces apoptotic death. Here, this possibility was tested by reducing the extracellular Cl concentration. Methods: 

Human lymphoid U937 and epithelial HeLa cells were provided for experiments after exposing to isotonic electrolyte solution which contains 146 or 1 mmCl. Measurements of mean cell volume, caspase-3 activity and cell viability were performed by a Coulter-type cell size analyzer, a fluorometric assay and a colorimetric assay, respectively. Results: 

After exposure to low Cl solution in which most chloride was replaced with aspartate, gluconate, phosphate or methanesulphonate, both U937 and HeLa cells exhibited, for up to 60 min, shrinkage to a level (90–80%) significantly lower than that in control high Cl solution. Reduction in cell viability started within 2 h and reached below 20% within 8 h after exposure to low Cl solution. The cell death was found to be associated with caspase-3 activation and DNA fragmentation. Conclusions: 

Exposure to isotonic low Cl solution induced sustained shrinkage and thereafter apoptotic death in U937 and HeLa cells. Thus, it is suggested that sustained cell shrinkage per se provides a sufficient condition for apoptosis induction.
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Keywords: apoptosis; apoptotic volume decrease; normotonic cell shrinkage

Document Type: Research Article

Publication date: 01 May 2006

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