Involvement of caspase-3 in apoptosis induced by Viscum album var. coloratum agglutinin in HL-60 cells.
A cytotoxic lectin (Viscum album L. coloratum agglutinin, VCA) from Korean mistletoe was isolated by affinity chromatography on Sepharose 4B immobilized with asialofetuin. In HL-60 cells, addition of VCA resulted in a dose- and time-dependent growth suppression, morphological changes of apoptotic nuclei, and DNA fragmentation characteristics of apoptosis. To investigate how caspase-3 activation during VCA-induced apoptosis induces cleavages of PARP, the expression of PARP and the pattern of caspase-3 activation in HL-60 cells were investigated. The native and processed PARP forms typically seen in apoptotic cells were observed, and a decrease in expression of the 32-kDa form of caspase-3 in a dose-dependent manner was observed. The VCA-induced apoptosis was significantly inhibited by a caspase-3 specific inhibitor, z-DEVD-FMK, and the PARP processing and caspase-3 activation were also inhibited by the inhibitor. A possible involvement of cell cycle arrest in VCA-induced apoptosis was investigated by flow cytometry and the results suggested that the apoptotic effect of VCA is not involved in the induction of cell cycle arrest.
Lyu
S Y
SY
College of Natural Sciences, Seoul Women's University, Korea.
Park
W B
WB
Choi
K H
KH
Kim
W H
WH
D017209:Apoptosis; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism*; D002453:Cell Cycle; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D018922:HL-60 Cells; D006801:Humans; D037102:Lectins / Q000494:pharmacology*; D014784:Mistletoe; D009842:Oligopeptides / Q000494:pharmacology; D037121:Plant Lectins; D028321:Plant Preparations; D010940:Plant Proteins; D010946:Plants, Medicinal; D054790:Ribosome Inactivating Proteins, Type 2; D012967:Sodium Dodecyl Sulfate; D014118:Toxins, Biological / Q000494:pharmacology*
D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors; D037102:Lectins; D009842:Oligopeptides; D037121:Plant Lectins; D028321:Plant Preparations; D010940:Plant Proteins; D054790:Ribosome Inactivating Proteins, Type 2; D014118:Toxins, Biological; C110772:benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone; C009674:ribosome inactivating protein, Viscum; D012967:Sodium Dodecyl Sulfate; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases