Cell swelling activates phospholipase A2 in Ehrlich ascites tumor cells.
Ehrlich ascites tumor cells, loaded with 3H-labeled arachidonic acid and 14C-labeled stearic acid for two hours, were washed and transferred to either isotonic or hypotonic media containing BSA to scavenge the labeled fatty acids released from the cells. During the first two minutes of hypo-osmotic exposure the rate of 3H-labeled arachidonic acid release is 3.3 times higher than that observed at normal osmolality. Cell swelling also causes an increase in the production of 14C-stearic acid-labeled lysophosphatidylcholine. This indicates that a phospholipase A2 is activated by cell swelling in the Ehrlich cells. Within the same time frame there is no swelling-induced increase in 14C-labeled stearic acid release nor in the synthesis of phosphatidyl 14C-butanol in the presence of 14C-butanol. Furthermore, U7312, an inhibitor of phospholipase C, does not affect the swelling induced release of 14C-labeled arachidonic acid. Taken together these results exclude involvement of phospholipase A1, C and D in the swelling-induced liberation of arachidonic acid. The swelling-induced release of 3H-labeled arachidonic acid from Ehrlich cells as well as the volume regulatory response are inhibited after preincubation with GDP beta S or with AACOCF3, an inhibitor of the 85 kDa, cytosolic phospholipase A2. Based on these results we propose that cell swelling activates a phospholipase A2--perhaps the cytosolic 85 kDa type--by a partly G-protein coupled process, and that this activation is essential for the subsequent volume regulatory response.
Thoroed
S M
SM
August Krogh Institute, University of Copenhagen, Denmark.
Lauritzen
L
L
Lambert
I H
IH
Hansen
H S
HS
Hoffmann
E K
EK
D016718:Arachidonic Acid / Q000378:metabolism*; D001201:Ascites / Q000378:metabolism; D002468:Cell Physiological Phenomena; D003600:Cytosol / Q000201:enzymology; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D007553:Isotope Labeling; D010739:Phospholipase D / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism*; D054466:Phospholipases A1; D054467:Phospholipases A2; D014316:Tritium; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000378:metabolism
D014316:Tritium; D016718:Arachidonic Acid; D006160:Guanosine Triphosphate; D010741:Phospholipases A; D054466:Phospholipases A1; D054467:Phospholipases A2; D010738:Type C Phospholipases; D010739:Phospholipase D