Cancer Res Treat.  2003 Aug;35(4):341-348.

TRAIL-Mediated Apoptosis in Human Liver Chang Cells

• Park C
• Hong SW
• Jin SH
• Kim NS
• Cho KH
• Cheon JH
• Ahn JY
• Yang JK
• Park R

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 AND METHODS
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.

Keyword

Tumor necrosis factor; Apoptosis; Caspase; 2-D; Hepatoma

MeSH Terms

Apoptosis*
Blotting, Western
Carcinoma, Hepatocellular
Caspase 3
Caspase 9
Cell Survival
DNA
DNA Fragmentation
Electrophoresis
Electrophoresis, Gel, Two-Dimensional
Humans*
Liver*
Peptide Hydrolases
Proteome
Rivers
Tumor Necrosis Factor-alpha
Caspase 3
Caspase 9
DNA
Peptide Hydrolases
Proteome
Tumor Necrosis Factor-alpha