TRAIL-Mediated Apoptosis in Human Liver Chang Cells / Journal of the Korean Cancer Association, 대한암학회지
Cancer Research and Treatment
;
: 341-348, 2003.
Article
in Korean
| WPRIM
| ID: wpr-226924
ABSTRACT
PURPOSE:
Tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL)/APO-2L is a member of the TNF family that can kill a wide variety of tumor cells, but not normal cells. This study was designed to investigate the down stream target proteins in TRAIL-mediated apoptosis of human liver, Chang cells. MATERIALS ANDMETHODS:
The expressions of DR4/DR5 in hepatoma cells, including Chang, HepG2 and Hep3B cells, were determined by RT-PCR. Cell viability was measured by MTT assay and apoptosis was assessed by DNA fragmentation assay. The catalytic activity of caspase- family proteases, including caspase-3 and -9, was tested by using fluorogenic biosubstrates. Expression of apoptotic mediators, including procaspase-3 and PARP proteins, was measured by Western blotting. The expression profile of proteins in Chang cells by using two-dimensional (2-D) gel electrophoresis and MALDI-TOF.RESULTS:
The results demonstrated that TRAIL (100 ng/ml) induced the apoptotic death of Chang cells, as characterized by the ladder-pattern fragmentation of genomic DNA. TRAIL increased the enzymatic activity of caspase- 3, corresponding to the time of appearance of cleaved PARP and caspase-9. In 2-D gel electrophoresis and MALDI- TOF analysis, the comparison of control versus apoptotic cells in the protein expressions revealed that signal intensity of 7 spots were decreased, whereas 6 spots were increased among 300 spots. These spots were resolved and identified as a protein information by MALDI-TOF.CONCLUSION:
We suggested that TRAIL induces the apoptotic death of Chang cells via proteome alterations inducing caspase cascade.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Peptide Hydrolases
/
DNA
/
Electrophoresis, Gel, Two-Dimensional
/
Cell Survival
/
Blotting, Western
/
Tumor Necrosis Factor-alpha
/
Apoptosis
/
Carcinoma, Hepatocellular
/
Proteome
/
Rivers
Type of study:
Prognostic study
Limits:
Humans
Language:
Korean
Journal:
Cancer Research and Treatment
Year:
2003
Type:
Article
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