Anti-Proliferative Effects of Human Anti-FZD7 Single Chain Antibodies on Colorectal Cancer Cells
Message:
Abstract:
Background
Colorectal cancer (CRC) accounts for the third most common cancer-related death in human. Wnt growth factors signaling have an important role in the CRCs’ development. Frizzled family receptor 7 (FZD7) belongs to the 10-member Frizzled (FZD) family of receptors and has an important role in triggering Wnt signaling. Antibody-therapy is an important class of modern medicine. Human single chain antibodies, which are composed of VH-Linker-VL are small antigen-binding units with unique properties, which overcome monoclonal antibodies and are applied for certain cancer immunotargeting approaches.
Objectives
This study aimed at determining the anti-tumor properties of 2 anti-FZD7 scFvs phage antibodies.
Methods
Two specific anti-FZD7 scFv antibodies were assessed for their anti-cancer effects. Flow cytometry was used to investigate the binding ability of scFvs to SW-480 colorectal cancer cell line, and SKBR3 cells, as the negative FZD7 cell line. Cell proliferation assay was performed to evaluate the growth inhibition effects of the different concentrations of anti-FZD7scFv phage antibodies (1000 to 4000 phage/cell). Mann-Whitney U test was used to compare percentages of cell growth between treated and untreated cells.
Results
Fluorescent activated cell sorting (FACS) analysis revealed that scFv I and II bound to 62.4% and 63.7% of SW-480 cells, respectively, whereas 6.4% and 8.3% bound to SKBR-3 cells, as the negative control. Results of MTT assays on SW-480 demonstrated growth inhibition of 53%, 63%, and 83% for scFv I (3000 phage/cell) after 24, 48, and 72 hours, respectively, while scFv II (4000 phage/cell) treatment showed growth inhibition of 33%, 47%, and 63% for the same period of time, respectively. No significant inhibitory effect for scFvs on the cell growth of SKBR-3 cells was observed (P
Conclusions
Targeted therapy using antibodies has been considered as a new cancer immunotherapy in the recent years. In this study we applied 2 anti-FZD7 antibodies to assess their effect on colorectal cancer cells. Results demonstrated that both scFvs bound to the SW-480 colorectal cancer cell expressing FZD7 receptor. Significant cell growth inhibition for both anti-Fdz7 scFvs after 72 hours of incubation with the optimum amounts of the antibodies was detected. The stronger potential of scFv I for inhibiting the growth of cancer cells was also obtained. The use of scFvs against colorectal cancer cells offers a therapeutic potential for these antibodies in colorectal cancer immunotherapy.
Language:
English
Published:
Shiraz Emedical Journal, Volume:18 Issue: 3, Mar 2017
Page:
5
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