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Cancer Research and Treatment > Volume 35(4); 2003 > Article
Cancer Research and Treatment 2003;35(4): 341-348. doi: https://doi.org/10.4143/crt.2003.35.4.341
TRAIL-Mediated Apoptosis in Human Liver Chang Cells
Channy Park, Sung Wook Hong, Sung Ho Jin, Nam Song Kim, Kyung Ho Cho, Jin Ho Cheon, Jae Yeon Ahn, Jung Ku Yang, Raekil Park
1Department of Microbiology, School of Medicine, WonkwangUniversity, Iksan, Jeonbuk, Korea. rkpark@wonkwang.ac.kr
2Department of Preventive Medicine, School of Medicine,Wonkwang University, Iksan, Jeonbuk, Korea.
3Professional Graduate School of Oriental Medicine, WonkwangUniversity, Iksan, Jeonbuk, Korea.
  Published online: August 31, 2003.
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.
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.
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.
We suggested that TRAIL induces the apoptotic death of Chang cells via proteome alterations inducing caspase cascade.
Key words: Tumor necrosis factor;Apoptosis;Caspase;2-D;Hepatoma
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