Leukemia inhibitory factor induces the 85-kDa cytosolic phospholipase A2 gene expression in cultured human bronchial epithelial cells.
Leukemia inhibitory factor (LIF) has become increasingly recognized as an important regulator of inflammation. This study is designed to determine whether LIF has an effect on arachidonate metabolism in human airway epithelial cells. LIF (100 ng/ml) induced a significantly increased release of prelabeled [3H] arachidonic acid (AA) from the human bronchial epithelial cell line (BEAS 2B cell) as well as from the primary cultures of human bronchial epithelial cells. Exposure of the LIF stimulated BEAS 2B cells to calcium ionophore A23187 (10(-5) M, 15 min) caused a further increase of [3H]AA release. To identify the role of cytosolic phospholipase A2 (cPLA2) in this upregulation of AA release, further experiments were performed to determine the expression of cPLA2 in the BEAS 2B cells. Immunoblot analysis indicated that LIF increased cPLA2 protein expression. Ribonuclease protection assay showed that LIF induced an increase of cPLA2 mRNA levels following 3 h to 24 h treatment. Nuclear run-on experiments suggested that LIF upregulated cPLA2 gene expression through post-translational regulation. These results demonstrate that LIF induces cPLA2 gene expression and modulates arachidonate metabolism in airway epithelial cells.
Ikezono
T
T
Critical Care Medicine Department, National Institutes of Health, Bethesda, MD 20892, USA.
Wu
T
T
Yao
X L
XL
Levine
S
S
Logun
C
C
Angus
C W
CW
Shelhamer
J H
JH
D016718:Arachidonic Acid / Q000032:analysis; D001980:Bronchi / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D002478:Cells, Cultured; D004848:Epithelium / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects; D006131:Growth Inhibitors / Q000494:pharmacology*; D006801:Humans; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D008222:Lymphokines / Q000494:pharmacology*; D010741:Phospholipases A / Q000096:biosynthesis* / Q000235:genetics; D054467:Phospholipases A2; D012333:RNA, Messenger / Q000096:biosynthesis; D018121:Receptors, Cytokine / Q000032:analysis; D053606:Receptors, OSM-LIF; D014158:Transcription, Genetic / Q000187:drug effects
D006131:Growth Inhibitors; D015850:Interleukin-6; C507837:LIF protein, human; C507840:LIFR protein, human; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D008222:Lymphokines; D012333:RNA, Messenger; D018121:Receptors, Cytokine; D053606:Receptors, OSM-LIF; D016718:Arachidonic Acid; D000001:Calcimycin; D010741:Phospholipases A; D054467:Phospholipases A2