Objectives:
Among the most promising antibody formats in terms of inhibiting
carcinogenesis are single-stranded variable fragments, whose targeted
binding to the Fzd7 receptor has been proven effective at suppressing
tumorigenesis. In this study, we investigated the effectiveness of an
anti-Fzd7 antibody fragment against both tumor growth and metastasis of
breast cancer cells.
Methods:
To develop anti-Fzd7 antibodies, bioinformatics approaches were
used and the antibodies were expressed recombinantly in E. coli BL21
(DE3). The expression of anti-Fzd7 fragments was verified by Western
blotting. Analysis of the antibodys binding capacity to Fzd7 was
conducted by flow cytometry. Cell death and apoptosis were assessed by
MTT and Annexin V/PI assays. The transwell migration and invasion
assays, as well as the scratch method, were used to evaluate cell
motility and invasiveness.
Results:
The anti-Fzd7 antibody was expressed successfully as a single band
of 31 kDa. It could bind to 21.5% of MDA-MB-231 cells, as opposed to
only 0.54% of SKBR-3 cells as negative control. According to MTT assay,
induced apoptosis was 73.7% in MDA-MB-231 cells, compared with 29.5% in
SKBR-3 cells. Also, the antibody exerted a significant inhibitory effect
of 76% and 58% on migration and invasion of MDA-MB-231 cells,
respectively.
Conclusion:
The recombinantly developed anti-Fzd7 scFv of this study could
exhibit significant antiproliferative and antimigratory properties,
along with a high apoptosis-inducing potential, making it suitable for
the immunotherapy of triple negative breast cancer.