Exp Mol Med.  1998 Mar;30(1):21-27.

Changes of phospholipase D activity in TNF-alpha and anti-Fas/Apo1 monoclonal antibody induced apoptosis in HL-60 and A20 cells

• Kang JH
• Shin IC
• Han JS

Abstract

The changes of phospholipase D (PLD) activity were investigated during the courses of apoptotic process induced by tumor necrosis factor (TNF)-alpha or anti-Fas/Apo1 antibody in human premyelocyte HL-60 and murine B cell lymphoma A20 cells. The treatment of recombinant TNF-alpha to HL-60 cells resulted in the increased PLD activity as determined by the phosphatidylethanol formation in the presence of 1% ethanol. The enhancement of PLD activity was also observed in the anti-Fas/Apo1 monoclonal antibody-treated A20 cells. However, the activity of PLD was maximized when HL-60 and A20 cells were treated with either TNF-alpha or anti-Fas/Apo1 monoclonal antibody for 6 h. Both TNF-alpha and anti-Fas/Apo1 monoclonal antibody increased PLD activity in a dose-dependent manner up to 200 U/ml and 200 ng/ml, respectively. When the intracellular activity of protein kinase C (PKC) was interrupted by treatment of calphostin-C, both the PLD activation and the apoptosis induced by TNF-alpha and anti-Fas/Apo1 monoclonal antibody appeared to be inhibited. Since PKC is reported to activate PLD, the
results
indicate that the intracellular signaling cascade via PLD may play a role in the induction of apoptosis induced by TNF-alpha and anti-Fas/Apo1 monoclonal antibody.

Keyword

apoptosis; anti-Fas monoclonal antibody; phospholipase D; protein kinase C; tumor necrosis factor-alpha

MeSH Terms

Animal
Antibodies, Monoclonal/pharmacology
Antigens, CD95/metabolism*
Antigens, CD95/immunology
Apoptosis*
DNA Fragmentation
Dose-Response Relationship, Drug
Enzyme Activation
HL-60 Cells
Human
Leukemia, Promyelocytic, Acute
Lymphoma, B-Cell
Mice
Naphthalenes/pharmacology
Phospholipase D/metabolism*
Protein Kinase C/antagonists & inhibitors
Receptors, Tumor Necrosis Factor/metabolism*
Signal Transduction
Tumor Necrosis Factor/pharmacology*