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Am J Physiol Lung Cell Mol Physiol 283: L573-L584, 2002. First published May 3, 2002; doi:10.1152/ajplung.00410.2001
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Vol. 283, Issue 3, L573-L584, September 2002

Elevated expression of hexokinase II protects human lung epithelial-like A549 cells against oxidative injury

Aftab Ahmad1, Shama Ahmad1, B. Kelly Schneider1, Corrie B. Allen1, Ling-Yi Chang2, and Carl W. White1

Departments of 1 Pediatrics and 2 Medicine, National Jewish Medical and Research Center, Denver, Colorado 80206

Increased glucose utilization and hexokinase (HK)-II expression are adaptive features of lung cells exposed to hypoxia or hyperoxia. HK-II is the most regulated isoform of HK. Whether its overexpression could be protective against oxidative stress was explored in human lung epithelial-like (A549) cells. HK-II was overexpressed in A549 cells in a tetracycline-repressible retroviral vector system. Elevated expression of HK-II was confirmed by Western blot and activity measurements. Cell death caused by exposure to hyperoxia was decreased in HK-II-overexpressing cells. This effect was reversed when HK-II expression was suppressed with doxycycline. A similar protective effect was observed in HK-II-overexpressing cells after treatment with 1 mM hydrogen peroxide for 48 h. At baseline, fluorescence microscopy showed that overexpressed HK-II was localized to mitochondria. Electron microscopic studies showed that hyperoxia-exposed HK-II overexpressors had better-preserved and quantitatively smaller mitochondria than those in which the HK-II expression was suppressed or in the nontransduced A549 cells. Mitochondrial membrane potential was increased in HK-II-overexpressing cells exposed to hyperoxia compared with the nontransduced control cells under similar conditions. The present study demonstrates that HK-II protects human lung epithelial-like A549 cells against oxidative insults by protecting the mitochondria.

hypoxia; hyperoxia; mitochondria


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