TRAIL-Mediated Apoptosis in Human Liver Chang Cells.
- Author:
Channy PARK
1
;
Sung Wook HONG
;
Sung Ho JIN
;
Nam Song KIM
;
Kyung Ho CHO
;
Jin Ho CHEON
;
Jae Yeon AHN
;
Jung Ku YANG
;
Raekil PARK
Author Information
1. Department of Microbiology, School of Medicine, Wonkwang University, Iksan, Jeonbuk, Korea. rkpark@wonkwang.ac.kr
- Publication Type:Original Article
- Keywords:
Tumor necrosis factor;
Apoptosis;
Caspase;
2-D;
Hepatoma
- MeSH:
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
- From:Cancer Research and Treatment
2003;35(4):341-348
- CountryRepublic of Korea
- Language:Korean
-
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.
