Caspase 3 expression is altered in a coculture model of neuroblastoma.
BACKGROUND
Previously, we demonstrated that neuroblastoma cells cocultured with hepatocytes are protected from apoptosis, while apoptosis is upregulated in the hepatocytes. The mechanisms responsible for these findings are unknown. We hypothesize that caspase 3, a cysteine protease central to the apoptotic pathway, will be altered in this coculture model that simulates metastatic neuroblastoma.


METHODS
Control human neuroblastoma cells and liver cells are plated in standard media. For the study group, a noncontact, coculture system is used. Hepatocytes are plated on cell culture inserts, placed above a growing layer of neuroblastoma cells, and incubated. Activated caspase 3 is measured after 1, 2, 3, or 4 days.


RESULTS
Activated caspase 3 levels are significantly decreased in the cocultured neuroblastoma cells on days 2, 3, and 4. Conversely, cocultured hepatocytes have a significant increase in caspase 3 activation at all time periods, with the largest difference seen after 1 day in coculture.


CONCLUSIONS
The capacity for neuroblastoma to differentially alter caspase 3 activation may provide a significant survival advantage for the neuroblastoma cells in metastatic environments. Understanding the mechanisms for this altered regulation may lead to improved and better targeted therapy for this malignancy.
Beierle
Elizabeth A
EA
Department of Surgery, University of Florida, Gainesville 32610, USA.
Dai
Wei
W
Langham
Max R
MR
Copeland
Edward M
EM
Chen
Mike K
MK
D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D018920:Coculture Techniques; D004789:Enzyme Activation; D022781:Hepatocytes / Q000201:enzymology; D006801:Humans; D009447:Neuroblastoma / Q000201:enzymology*; D013997:Time Factors; D014407:Tumor Cells, Cultured
C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases