Betulinic acid and its derivatives have been extensively studied in the past for their anti-tumor effects, but relatively little is known about its precursor betulin. In this study we showed that betulin, an abundant natural product, significantly inhibits the cell growth of human hepatoma HepG2 cells in a dose-dependent manner. In the presence of 10 μg/mL betulin, HepG2 cells undergo an apoptosis, as evidenced by apoptotic morphology such as cell shrinkage, membrane blebbing, nuclear condensation and fragmentation, apoptotic body formation, and caspase activation. Kinetics analysis shows that the depolarization of the mitochondrial membrane potential and the release of the mito- chondrial apoptotic protein cytochrome c occurred as early as 2 h post treatment of HepG2 cells with 10 μg/mL betulin. Proteolytic activation of caspase-9, but not caspase-8, was observed in this apoptosis process. Moreover, the inactivation of caspase-9 by its specific siRNA dramatically reduced betulin-induced caspase-3 activation and apoptosis. Taken together, our observations indicate that the activation of caspase-9 is critical for betulin-induced apoptosis of human hepatoma HepG2 cells.
LI YangSHEN Jared TaiyiGAO ChangLI QingJIN Ying-hua
Recent observations indicate that the resistance of apoptosis is an important process of tumor metastasis and metastases are the cause of 90% of human cancer death. Etoposide, a semisynthetic derivative of the podophyllotoxins, is a clinically used anti-cancer reagent, but the effects of it on metastatic gastric carcinoma cells are totally unknown. In this study, etoposide induced apoptotic cell death in human gastric adenocarcinoma cell line SGC-7901, derived from metastatic lymph nodes, as evidenced by the analysis of DNA fragmentation, apoptotic body formation, caspase activation, and apoptosis specific changes in cell morphology is demonstrated. The depolarization of mitochondrial membrane and the release of cytochrome c were most early events in etoposide treated SGC-7901 cells, and were followed by caspase-3 activation and PARP cleavage. Caspase-8 activation was not detected under the same condition. Thus, it was proposed that etoposide induces caspase-associated apoptotic cell death in human metastatic gastric carcinoma, which is initiated by mitochondrial cytochrome c release.
LI Jing-huaCHEN YueWANG Jia-siKONG WeiJIN Ying-hua
