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Am J Physiol Lung Cell Mol Physiol (May 24, 2002). doi:10.1152/ajplung.00025.2002
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Articles in PresS, published online ahead of print May 24, 2002
Am J Physiol Lung Cell Mol Physiol, 10.1152/ajplung.00025.2002
Submitted on January 18, 2002
Accepted on April 27, 2002

A549 subclones demonstrate heterogeneity in toxicological sensitivity and antioxidant profile

Nobuo Watanabe1, Dale A. Dickinson1, David M. Krzywanski1, Karen E. Iles1, Hongqiao Zhang1, Charles J. Venglarik1, and Henry J. Forman1*

1 Department of Environmental Health Sciences, University of Alabama at Birmingham, Birmingham, AL, USA

* To whom correspondence should be addressed. E-mail: hforman{at}uab.edu.

In A549 cell culture, significant variability was found in sensitivity to actinomycin D. Using limiting dilution, actinomycin D-susceptible (G4S) and resistant (D3R) subclones were isolated. G4S cells were also susceptible to protein synthesis inhibitors, a redox cycling quinone, and an electrophile, with concomitant activation of caspases 3 and 9. D3R cells were resistant to these agents, without caspase activation. Antioxidant profiles revealed that D3R cells had significantly higher glutathione and glutathione reductase activity but markedly lower catalase, glutathione peroxidase and aldehyde reductase activities than G4S cells. Thus, A549 cells contain at least two distinct subpopulations with respect to predisposition to cell death and antioxidant profile. As sensitivities to agents and the antioxidant profile were inconsistent, mechanisms independent of antioxidants, including the apparent inability to activate caspases in D3R cells, may play an important role. Regardless, the results suggest that antioxidant profiles of asymmetrical cell populations cannot predict sensitivity to oxidants, and warn that the use of single subclones is advisable for mechanistic studies using A549 or other unstable cell lines.




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