TRAIL-Mediated Apoptosis in Human Liver Chang Cells

• Published in: Cancer research and treatment Vol. 35; no. 4; pp. 341 - 348
• Main Authors: Park, Channy, Hong, Sung Wook, Jin, Sung Ho, Kim, Nam Song, Cho, Kyung Ho, Cheon, Jin Ho, Ahn, Jae Yeon, Yang, Jung Ku, Park, Raekil
• Format: Journal Article
• Language: English
• Published: Korea (South) 대한암학회 01.08.2003
• Subjects: 의약학; Caspase; 2-D; Hepatoma; Tumor necrosis factor; Apoptosis
• ISSN: 1598-2998; 2005-9256
• DOI: 10.4143/crt.2003.35.4.341

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.