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Chemosensitivity testing of oral cancer cells treated with a p185neu-specific agent
R Werkmeister1, T Fillies, B Brandt
1Department of Oral and Maxillofacial Surgery, University of Münster, Germany. werkmei@uni-muenster.de
Abstract:
The amplification and overexpression of the erbB-2 oncogene and its involvement in tumorigenesis makes this receptor an appropriate target for specific agents directed towards tumor cells. The purpose of this study was to evaluate the in vitro effect of the bacterially produced recombinant immunotoxin scFv(FRP5)-ETA on the protein synthesis and adenosine triphosphate (ATP) reduction in oral squamous cell carcinoma (OSCC) cells. This agent recognizes the erbB-2 receptor and inhibits protein synthesis in receptor-overexpressing cells. OSCC cells were selected for this study, and amplification and expression levels of the erbB-2 receptor were determined. Cell suspensions were cultured for 6 d with various concentrations of scFv(FRP5)-ETA (1-1000 ng/ml). A431 and MDA-MB468 cell lines were used as controls. Chemosensibility of tumor cells was measured by [3H]leucine incorporation assay and by an ATP luminescence assay. In OSCC cells with amplification and overexpression of erbB-2 inhibition, up to 92% of protein synthesis and 90% of ATP reduction was observed when cells were exposed to 1,000 ng/ml immunotoxin. In OSCC cells showing a deletion of erbB-2 and in erbB-2-negative MDA-MB468 cells, protein synthesis was inhibited by 22% and 8%, respectively. These results indicate that the effectiveness of a recombinant immunotoxin targeting erbB-2 receptors in OSCC cells depends on the level of erbB-2 amplification and expression, that it is highly specific for tumor cells expressing these receptors, and that a dose-dependency can be observed.
Insights
This study shows that a targeted immunotoxin effectively reduces protein synthesis and ATP levels in oral squamous cell carcinoma (OSCC) cells overexpressing the erbB-2 receptor. Its efficacy is dependent on erbB-2 expression levels, demonstrating high specificity for tumor cells.
Area of Science:
- Oncology
- Molecular Biology
- Biotechnology
Background:
- The erbB-2 oncogene's amplification and overexpression are implicated in tumorigenesis.
- Targeting erbB-2 is a promising strategy for cancer therapy.
- Oral squamous cell carcinoma (OSCC) cells often exhibit erbB-2 alterations.
Purpose of the Study:
- To evaluate the in vitro efficacy of a recombinant immunotoxin, scFv(FRP5)-ETA, against OSCC cells.
- To assess the immunotoxin's impact on protein synthesis and adenosine triphosphate (ATP) levels.
- To determine the correlation between erbB-2 expression and immunotoxin effectiveness.
Main Methods:
- OSCC cell lines with varying erbB-2 amplification and expression levels were cultured.
- Cells were treated with a range of scFv(FRP5)-ETA concentrations (1-1000 ng/ml).
- Protein synthesis was measured using [3H]leucine incorporation, and cell viability via ATP luminescence assay.
Main Results:
- A significant inhibition of protein synthesis (up to 92%) and ATP reduction (up to 90%) was observed in OSCC cells with high erbB-2 expression when treated with 1000 ng/ml immunotoxin.
- Minimal inhibition (22% and 8%) occurred in OSCC cells with erbB-2 deletion or negative expression, respectively.
- Results demonstrated a clear dose-dependent and erbB-2-specific effect of the immunotoxin.
Conclusions:
- The recombinant immunotoxin scFv(FRP5)-ETA is effective against OSCC cells.
- Immunotoxin efficacy is directly related to the level of erbB-2 amplification and expression.
- The agent exhibits high specificity for erbB-2-expressing tumor cells, supporting its therapeutic potential.
