The
anti-apoptotic protein Bcl-2 is upregulated in several
cancers, including
diffuse large B-cell lymphoma (DLBCL) and
chronic lymphocytic leukemia (CLL). In a subset of these
cancer cells, Bcl-2 blocks Ca2+-mediated apoptosis by suppressing the function of
inositol 1,4,5-trisphosphate (IP3) receptors (IP3Rs) located at the endoplasmic reticulum (ER). A
peptide tool, called Bcl-2/
IP3 receptor disruptor-2 (BIRD-2), was developed to disrupt Bcl-2/IP3R complexes, triggering pro-apoptotic Ca2+ signals and killing Bcl-2-dependent
cancer cells. In DLBCL cells, BIRD-2 sensitivity depended on the expression level of IP3R2 channels and constitutive IP3 signaling downstream of the B-cell receptor. However, other cellular pathways probably also contribute to BIRD-2-provoked cell death. Here, we examined whether BIRD-2-induced apoptosis depended on extracellular Ca2+ and more particularly on store-operated Ca2+ entry (SOCE), a Ca2+-influx pathway activated upon ER-store depletion. Excitingly,
DPB162-AE, a SOCE inhibitor, suppressed BIRD-2-induced cell death in DLBCL cells. However,
DPB162-AE not only inhibits SOCE but also depletes the ER Ca2+ store. Treatment of the cells with
YM-58483 and
GSK-7975A, two selective SOCE inhibitors, did not protect against BIRD-2-induced apoptosis. Similar data were obtained by knocking down STIM1 using
small interfering RNA. Yet, extracellular Ca2+ contributed to BIRD-2 sensitivity in DLBCL, since the extracellular Ca2+ buffer
ethylene glycol tetraacetic acid (
EGTA) blunted BIRD-2-triggered apoptosis. The protective effects observed with
DPB162-AE are likely due to ER Ca2+-store depletion, since a similar protective effect could be obtained using the
sarco/endoplasmic reticulum Ca2+-ATPase inhibitor
thapsigargin. Thus, both the ER Ca2+-store content and extracellular Ca2+, but not SOCE, are critical factors underlying BIRD-2-provoked cell death.