Related Experiment Videos
Role of amyloid precursor protein (APP): study with antisense transfection of human neuroblastoma cells
A C LeBlanc1, D M Kovacs, H Y Chen
1Department of Pathology, Case Western Reserve University, Cleveland, Ohio 44106.
Abstract:
The function of amyloid precursor protein (APP) was investigated in human neuroblastoma La-N-1 cells by stable transfection with a DNA construct encoding antisense APP mRNA. Levels of APP mRNA, as well as proteins, were reduced by 80-90% in antisense APP transfected (ASAT) cells. ASAT cells exhibited three main features as a result of APP gene expression deprivation: (1) a 30% reduction in cell proliferation, (2) reduced cell adhesion that could be reversed by the addition of La-N-1 conditioned media as a source of secreted APP, and (3) a two- and four-fold increase in neurite-bearing cells suggesting that cellular APP may be involved in neurite extension. The first two features confirm previously reported functions for APP in proliferation and adhesion of non-neuronal cell types but the use of neuroblastoma cells in this study disclose a novel role for cellular APP in neurite extension.
Insights
Amyloid precursor protein (APP) deprivation in neuroblastoma cells reduced proliferation and adhesion, confirming known functions. This study reveals a novel role for cellular APP in promoting neurite extension.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- Amyloid precursor protein (APP) is implicated in various cellular functions.
- Previous research has highlighted APP's role in non-neuronal cell proliferation and adhesion.
Purpose of the Study:
- To investigate the function of amyloid precursor protein (APP) in human neuroblastoma La-N-1 cells.
- To elucidate the role of cellular APP in neuroblastoma cell behavior, including proliferation, adhesion, and neurite extension.
Main Methods:
- Stable transfection of La-N-1 cells with antisense APP mRNA to create antisense APP transfected (ASAT) cells.
- Quantification of APP mRNA and protein levels, cell proliferation rates, cell adhesion, and neurite extension in ASAT cells compared to controls.
- Assessment of the effect of secreted APP on cell adhesion in ASAT cells.
Main Results:
- ASAT cells showed an 80-90% reduction in APP mRNA and protein levels.
- A 30% decrease in cell proliferation and reduced cell adhesion were observed in ASAT cells.
- A significant increase in neurite-bearing cells (two- and four-fold) was noted in ASAT cells, suggesting a role for cellular APP in neurite extension.
Conclusions:
- Cellular APP plays a role in regulating neuroblastoma cell proliferation and adhesion.
- This study identifies a novel function for cellular APP in promoting neurite extension in neuroblastoma cells.
- The findings suggest that APP's role extends beyond its known functions in non-neuronal cells, with implications for neuronal development.