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Truncated DCC reduces N-cadherin/catenin expression and calcium-dependent cell adhesion in neuroblastoma cells
M Reyes-Múgica1, J A Meyerhardt, J Rzasa
1Department of Pathology, Yale University School of Medicine and West Haven Veterans Administration Medical Center, New Haven, Connecticut 06520-8023,USA. miguel.reyes@yale.edu
Abstract:
The deleted in colorectal cancer (DCC) protein is important in the pathway guidance of cells and cell processes during neural development, and DCC has also been implicated in the aberrant cellular migrations of neuroblastoma dissemination. We attempted to further define DCC protein function by the overexpression of full-length and truncated DCC constructs in a human neuroblastoma cell line. Overexpression of the truncated DCC protein resulted in a less epithelioid morphology. This was accompanied by decreases in expression of N-cadherin and alpha- and beta-catenin by immunoblot and Northern blot analysis. Levels of desmoglein were relatively less affected, whereas endogenous DCC protein levels were increased in the truncated transfectants. N-cadherin immunofluorescence was consistent with the immunoblot studies and localized the protein to the cytoplasm and sites of cell-cell contact. Cell aggregation studies demonstrated diminished calcium-dependent aggregation in the truncated transfectants. In conclusion, overexpression of a truncated DCC protein in neuroblastoma cells resulted in the loss of an epithelioid morphology, diminished expression of N-cadherin and alpha- and beta-catenin, and diminished calcium-dependent cell adhesion. These studies provide the first evidence of an apparent functional link between DCC and N-cadherin/catenin-dependent cell adhesion.
Insights
Overexpressing a truncated Deleted in Colorectal Cancer (DCC) protein in neuroblastoma cells reduced their epithelioid shape and cell adhesion. This study links DCC to N-cadherin and catenin-dependent cell adhesion mechanisms.
Area of Science:
- Cell Biology
- Neuroscience
- Cancer Research
Background:
- The Deleted in Colorectal Cancer (DCC) protein is crucial for neural development and implicated in neuroblastoma cell migration.
- Understanding DCC protein function is vital for neuroblastoma research.
Purpose of the Study:
- To investigate the function of DCC protein by overexpressing full-length and truncated DCC constructs in a human neuroblastoma cell line.
- To define the role of DCC in neuroblastoma cell morphology and adhesion.
Main Methods:
- Overexpression of full-length and truncated DCC constructs in human neuroblastoma cells.
- Immunoblot and Northern blot analysis to assess protein and gene expression (N-cadherin, alpha-catenin, beta-catenin, desmoglein).
- Immunofluorescence for N-cadherin localization and cell aggregation studies for cell adhesion analysis.
Main Results:
- Overexpression of truncated DCC led to a less epithelioid morphology.
- Decreased expression of N-cadherin and alpha-/beta-catenin was observed.
- Diminished calcium-dependent cell aggregation was evident in truncated DCC transfectants.
Conclusions:
- Truncated DCC protein overexpression in neuroblastoma cells causes loss of epithelioid morphology and reduced cell adhesion.
- This study establishes a functional link between DCC and N-cadherin/catenin-dependent cell adhesion.
- Findings provide insights into DCC's role in neuroblastoma cell behavior and potential therapeutic targets.