ARACHIDONATE AND OTHER FATTY-ACIDS MOBILIZE CA2-GLUCURONIDASE RELEASEIN A CA2+-DEPENDENT FASHION FROM UNDIFFERENTIATED HL-60 CELLS( IONS AND STIMULATE BETA)
De. Packham et al., ARACHIDONATE AND OTHER FATTY-ACIDS MOBILIZE CA2-GLUCURONIDASE RELEASEIN A CA2+-DEPENDENT FASHION FROM UNDIFFERENTIATED HL-60 CELLS( IONS AND STIMULATE BETA), Cell calcium, 17(6), 1995, pp. 399-408
Arachidonate (1-300 mu M) mobilized Ca2+ ions from an intracellular st
ore and stimulated the entry of Ca2+ ions from the extracellular fluid
in undifferentiated HL-60 cells that had been loaded with Fura-2. The
integrated response was biphasic in farm: arachidonate liberated Ca2 ions from the intracellular store first, resulting in a transient inc
rease in cytosolic free Ca2+ concentration ([Ca2+](i)). Ca2+ entry fro
m the extracellular fluid was not evident for a further 1-2 min, At ba
seline, [Ca2+](i) was 48.1 +/- 14.0 nM (SEM, n = 5), Upon addition of
arachidonate (100 mu M), [Ca2+](i) rose to a transient peak level of 2
17 +/- 38.6 nM (SEM, n = 5) and a later plateau level of 427 +/- 118 n
M (SEM, n = 5). Removal of added Ca2+ ions from the extracellular flui
d in the presence of EGTA (1.0 mM) had no effect on the initial transi
ent response but abolished the second phase of the response, In HL-60
cells that had been loaded with BAPTA/AM, the initial transient phase
of the response was abolished but the elevation in [Ca2+](i) due to Ca
2+ entry from the extracellular fluid was unaffected. Undifferentiated
HL-60 cells also responded to arachidonate (100 mu M) with an increas
e in the release of the lysosomal enzyme beta-glucuronidase. Arachidon
ate-induced beta-glucuronidase release from BAPTA-loaded cells or in c
ontrol cells exposed to Ca2+-free solutions was inhibited by about 50%
. In BAPTA-loaded cells that were incubated with Ca2+-free solutions,
arachidonate-induced beta-glucuronidase release was inhibited by about
90%. Leukotriene B-4 failed to elevate [Ca2+](i) in the concentration
range 0.01-1 mu M and failed to activate beta-glucuronidase release i
n concentrations up to 10 mu M. Furthermore, the cyclo-oxygenase/lipox
ygenase inhibitor ETYA (100 mu M) was without effect on secretion. Con
sistent with this finding, we found that a large number- of unsaturate
d fatty acids could reproduce the effect of arachidonate on [Ca2+](i)
and beta-glucuronidase release. Fatty acids belonging to the omega-3,
omega-6 and omega-7 unsaturated fatty acid families were effective in
elevating [Ca2+](i) and stimulating beta-glucuronidase release. Howeve
r, three unsaturated fatty acids, all belonging to the omega-9 fatty a
cid family, were ineffective.