Acquired resistance of a mammalian cell line to hypoxia-reoxygenation through cotransfection of Kir6.2 and SUR1 clones.

Reoxygenation after transient hypoxia is a common clinical condition that often causes greater tissue damage than persistent hypoxia itself. This warrants the development of a means to protect cells against hypoxia-reoxygenation injury. Adenosine triphosphate (ATP)-sensitive K+ (KATP) channels have been proposed to play an essential role in the mechanisms of endogenous cellular protection. Thus far, however, KATP channel proteins have not been exploited to generate an injury-resistant cellular phenotype by delivering KATP channel genes into injury-prone cells. A first step in this direction is the evaluation of the outcome of transferring genes encoding KATP channels into a KATP channel-deficient cell type exposed to metabolic stress. Untransfected COS-7 monkey kidney cells, which natively lack KATP channels, were found to be vulnerable to hypoxia-reoxygenation injury, which induced cytosolic Ca2+ loading, as measured by digital epifluorescent imaging. COS-7 cells cotransfected with KATP channel genes, Kir6.2 and SUR1, gained resistance to hypoxia-reoxygenation. This acquired resistance was abolished by glyburide, the KATP channel antagonist. We have previously shown that Kir6.2 and SUR1 physically associate to form a functional KATP channel, not reconstituted by either of the subunits alone. Transfection with individual channel subunits, Kir6.2 or SUR1, failed to produce resistance to hypoxia-reoxygenation induced Ca2+ loading. This is a first demonstration that transfer of KATP channel subunits can generate an injury-resistant cellular phenotype. The findings from this study may, thus, provide a framework for future therapeutic strategies based on gene delivery of KATP channel subunits in cells and tissues vulnerable to hypoxia-reoxygenation insults.