TY - JOUR
T1 - CD40 induces human multiple myeloma cell migration via phosphatidylinositol 3-kinase/AKT/NF-κB signaling
AU - Tai, Yu-Tzu
AU - Podar, Klaus
AU - Mitsiades, Nicholas
AU - Lin, Boris
AU - Mitsiades, Constantine
AU - Gupta, Deepak
AU - Akiyama, Masaharu
AU - Catley, Laurence
AU - Hideshima, Teru
AU - Munshi, Nikhil C
AU - Treon, Steven P
AU - Anderson, Kenneth C
PY - 2003/4/1
Y1 - 2003/4/1
N2 - Multiple myeloma (MM) is characterized by clonal expansion of malignant plasma cells in the bone marrow and their egress into peripheral blood with progression to plasma cell leukemia. Our previous study defined a functional role of CD40 activation in MM cell homing and migration. In this study, we examine signaling events mediating CD40-induced MM cell migration. We show that cross-linking CD40, using either soluble CD40L (sCD40L) or anti-CD40 monoclonal antibody (mAb), induces phosphatidylinositol 3-kinase (PI3K) activity and activates its downstream effector AKT in MM.1S cells. CD40 activation also activates the MAP kinase (MEK) pathway, evidenced by phosphorylation of extracellular signal-regulated mitogen-activated protein kinase (ERK), but not c-jun amino-terminal kinase (JNK) or p38, in a dose- and time-dependent manner. Using pharmacologic inhibitors of PI3K and MEK, as well as adenoviruses expressing dominant-negative and constitutively expressed AKT, we demonstrate that PI3K and AKT activities are required for CD40-induced MM cell migration. In contrast, inhibition of ERK/MEK phosphorylation only partially (10%-15%) prevents migration, suggesting only a minor role in regulation of CD40-mediated MM migration. We further demonstrate that CD40 induces nuclear factor (NF)-κB activation as a downstream target of PI3K/AKT signaling, and that inhibition of NF-κB signaling using specific inhibitors PS1145 and SN50 completely abrogates CD40-induced MM migration. Finally, we demonstrate that urokinase plasminogen activator (uPA), an NF-κB target gene, is induced by CD40; and conversely, that uPA induction via CD40 is blocked by PI3K and NF-κB inhibitors. Our data therefore indicate that CD40-induced MM cell migration is primarily mediated via activation of PI3K/AKT/NF-κB signaling, and further suggest that novel therapies targeting this pathway may inhibit MM cell migration associated with progressive MM.
AB - Multiple myeloma (MM) is characterized by clonal expansion of malignant plasma cells in the bone marrow and their egress into peripheral blood with progression to plasma cell leukemia. Our previous study defined a functional role of CD40 activation in MM cell homing and migration. In this study, we examine signaling events mediating CD40-induced MM cell migration. We show that cross-linking CD40, using either soluble CD40L (sCD40L) or anti-CD40 monoclonal antibody (mAb), induces phosphatidylinositol 3-kinase (PI3K) activity and activates its downstream effector AKT in MM.1S cells. CD40 activation also activates the MAP kinase (MEK) pathway, evidenced by phosphorylation of extracellular signal-regulated mitogen-activated protein kinase (ERK), but not c-jun amino-terminal kinase (JNK) or p38, in a dose- and time-dependent manner. Using pharmacologic inhibitors of PI3K and MEK, as well as adenoviruses expressing dominant-negative and constitutively expressed AKT, we demonstrate that PI3K and AKT activities are required for CD40-induced MM cell migration. In contrast, inhibition of ERK/MEK phosphorylation only partially (10%-15%) prevents migration, suggesting only a minor role in regulation of CD40-mediated MM migration. We further demonstrate that CD40 induces nuclear factor (NF)-κB activation as a downstream target of PI3K/AKT signaling, and that inhibition of NF-κB signaling using specific inhibitors PS1145 and SN50 completely abrogates CD40-induced MM migration. Finally, we demonstrate that urokinase plasminogen activator (uPA), an NF-κB target gene, is induced by CD40; and conversely, that uPA induction via CD40 is blocked by PI3K and NF-κB inhibitors. Our data therefore indicate that CD40-induced MM cell migration is primarily mediated via activation of PI3K/AKT/NF-κB signaling, and further suggest that novel therapies targeting this pathway may inhibit MM cell migration associated with progressive MM.
KW - CD40 Antigens/metabolism
KW - Cell Movement
KW - Humans
KW - Ligands
KW - Mitogen-Activated Protein Kinases/metabolism
KW - Multiple Myeloma/pathology
KW - NF-kappa B/metabolism
KW - Phosphatidylinositol 3-Kinases/metabolism
KW - Phosphorylation
KW - Protein Serine-Threonine Kinases
KW - Proto-Oncogene Proteins/metabolism
KW - Proto-Oncogene Proteins c-akt
KW - Signal Transduction
KW - Tumor Cells, Cultured
KW - Urokinase-Type Plasminogen Activator/metabolism
UR - http://www.scopus.com/inward/record.url?scp=0038784379&partnerID=8YFLogxK
U2 - 10.1182/blood-2002-09-2813
DO - 10.1182/blood-2002-09-2813
M3 - Journal article
C2 - 12433678
SN - 0006-4971
VL - 101
SP - 2762
EP - 2769
JO - Blood
JF - Blood
IS - 7
ER -