Betulinic acid induces apoptosis in human neuroblastoma cell lines
M L Schmidt1, K L Kuzmanoff, L Ling-Indeck
1Department of Pediatrics, College of Medicine, University of Illinois at Chicago 60612-7324, USA.
Abstract:
Neuroblastoma has long been recognized to show spontaneous regression during fetal development and in the majority of stage 4s infants < 1 year of age with disseminated disease. Stage 4s disease regresses with no chemotherapy in 50% of the patients. The mechanism by which this occurs is not understood but may be programmed cell death or apoptosis. Betulinic acid (BA) has been reported to induce apoptosis in human melanoma with in vitro and in vivo model systems. Melanoma, like neuroblastoma, is derived from the neural crest cell. We hypothesised that neuroblastoma cells have the machinery for programmed cell death and that apoptosis could be induced by betulinic acid. Nine human neuroblastoma cell lines were treated in vitro with BA at concentrations of 0-20 micrograms/ml for 0-6 days. Profound morphological changes were noted within 3 days. Cells withdrew their axonic-like extensions, became non-adherent and condensed into irregular dense spheroids typical of apoptotic cell death (ED50 = 14-17 micrograms/ml). DNA fragmentation analysis showed ladder formation in the 100-1200 bp region in 3/3 neuroblastoma cell lines treated with BA for 24-72 h. Thus, apparently BA does induce AP in neuroblastoma in vitro. This model will be utilised to investigate the role of apoptosis-related genes in neuroblastoma proliferation and to determine the therapeutic efficacy of BA in neuroblastoma in vivo.
Insights
Betulinic acid induces apoptosis in neuroblastoma cells, a programmed cell death pathway. This finding offers potential for new neuroblastoma treatments targeting this mechanism.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Neuroblastoma, a pediatric cancer, exhibits spontaneous regression in some cases.
- The mechanism of spontaneous regression, potentially involving apoptosis, remains unclear.
- Betulinic acid (BA) is known to induce apoptosis in melanoma, a cancer of neural crest origin.
Purpose of the Study:
- To investigate if betulinic acid can induce apoptosis in neuroblastoma cells.
- To explore the potential of BA as a therapeutic agent for neuroblastoma.
Main Methods:
- Treatment of nine human neuroblastoma cell lines with betulinic acid in vitro.
- Morphological assessment for signs of apoptotic cell death.
- DNA fragmentation analysis to confirm apoptosis.
Main Results:
- Betulinic acid induced significant morphological changes indicative of apoptosis in neuroblastoma cells.
- DNA fragmentation analysis confirmed apoptosis induction by BA.
- Effective concentration (ED50) for BA-induced apoptosis was determined.
Conclusions:
- Betulinic acid effectively induces apoptosis in human neuroblastoma cells in vitro.
- This provides a model for further research into apoptosis-related genes in neuroblastoma.
- Betulinic acid shows promise for in vivo therapeutic efficacy studies in neuroblastoma.
