논문검색
Author Channy Park1,3, Sung-Wook Hong2, Sung-Ho Jin2, Nam-song Kim2, Kyung-Ho Cho2, Jin-Ho Cheon2, Jae-Yeon Ahn2, Jung-ku Yang2 and Raekil Park1
Place of duty Departments of 1Microbiology and 2Preventive Medicine, School of Medicine; 3Professional Graduate School of Oriental Medicine, Wonkwang University, Iksan, Jeonbuk, Korea
Title TRAIL-Mediated Apoptosis in Human Liver Chang Cells
Publicationinfo Cancer Research and Treatment 2003 Aug; 035(04): 341-349.
Key_word Tumor necrosis factor, Apoptosis, Caspase, 2-D, Hepatoma
Full-Text
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. (Cancer Research and Treatment 2003;35:341-348