Mitochondrial but not plasmalemmal BK channels are hypoxia-sensitive in human glioma.
Tumor cells are resistant to hypoxia but the underlying mechanism(s) of this tolerance remain poorly understood. In healthy brain cells, plasmalemmal Ca(2+)-activated K(+) channels ((plasma)BK) function as oxygen sensors and close under hypoxic conditions. Similarly, BK channels in the mitochondrial inner membrane ((mito)BK) are also hypoxia sensitive and regulate reactive oxygen species production and also permeability transition pore formation. Both channel populations are therefore well situated to mediate cellular responses to hypoxia. In tumors, BK channel expression increases with malignancy, suggesting these channels contribute to tumor growth; therefore, we hypothesized that the sensitivity of (plasma)BK and/or (mito)BK to hypoxia differs between glioma and healthy brain cells. To test this, we examined the electrophysiological properties of (plasma)BK and (mito)BK from a human glioma cell line during normoxia and hypoxia. We observed single channel activities in whole cells and isolated mitoplasts with slope conductance of 199 ± 8 and 278 ± 10 pA, respectively. These currents were Ca(2+)- and voltage-dependent, and were inhibited by the BK channel antagonist charybdotoxin (0.1 μM). (plasma)BK could only be activated at membrane potentials >+40 mV and had a low open probability (NPo) that was unchanged by hypoxia. Conversely, (mito)BK were active across a range of membrane potentials (-40 to +40 mV) and their NPo increased during hypoxia. Activating (plasma)BK, but not (mito)BK induced cell death and this effect was enhanced during hypoxia. We conclude that unlike in healthy brain cells, glioma (mito)BK channels, but not (plasma)BK channels are oxygen sensitive.
Section of Respiratory Medicine, Department of Pediatrics, University of California San Diego, La Jolla, California.
Xiang Q
XQ
Gu
Matthew E
ME
Pamenter
Detlef
D
Siemen
Xiaolu
X
Sun
Gabriel G
GG
Haddad
D000255:Adenosine Triphosphate / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D018999:Charybdotoxin / Q000494:pharmacology; D005910:Glioma / Q000473:pathology / Q000648:ultrastructure*; D006801:Humans; D000860:Hypoxia / Q000378:metabolism*; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008928:Mitochondria / Q000378:metabolism* / Q000648:ultrastructure; D009498:Neurotoxins / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D011419:Propidium / Q000378:metabolism
D051036:Large-Conductance Calcium-Activated Potassium Channels; D009498:Neurotoxins; D018999:Charybdotoxin; D011419:Propidium; D000255:Adenosine Triphosphate; D002118:Calcium
cancer; electrophysiology; mitoplast patch