Reem Mebed , Yasser BM Ali, Nahla Shehata, Nadia El-Guendy, Nahla Gamal, Abdel-Rahman Zekri and Salwa Sabet*
Background: Bevacizumab (Bev) resistance is hypothesized to be overcome by combination with inhibitors of other signalling pathways.
Objective: We aimed to study the effect of combining Bev with knocked down β-catenin (Bev-β-cat-siRNA) on the expression of VEGF-A, Slug, NFКB and its two target genes c-Flip and FasR in HepG2. Expression of VEGF-A and Slug was also studied in Caco-2 cells.
Methods: Cultured cells were divided into six groups 1) cells treated with Bev only 2) cells treated with β-catenin-siRNA 3) cells treated with Bev-β-cat-siRNA 4) cells treated with negative control 5) cells treated with Bev-negative control and untreated cells. Expressions were assessed using qPCR and western blotting.
Results: Bev-β-cat-siRNA significantly reduced the mRNA level of VEGF-A, which was initially increased in response to Bev alone in HepG2 but not in Caco-2. Additionally, Bev-β-cat-siRNA significantly decreased Slug mRNA level compared to Bev only treated HepG2 cells. In contrast, VEGF-A and Slug mRNA levels in Bev only group were remarkably lower than Bev-β-cat-siRNA in Caco-2 cells. Distinct β-catenin and Slug protein expressions were noticed in HepG2 and Caco-2 cells. On the other hand, Bev-β-cat-siRNA remarkably reduced the level of NFКB, FasR and c-Flip compared to Bev only treated HepG2 cells although the difference was not statistically significant.
Conclusion: We conclude that, combining Bevacizumab with knocked down β-catenin reduce the expression of VEGF-A and Slug in HepG2 but not in Caco-2 cells.
Bevacizumab, β-catenin, VEGF-A, Slug, NFКB, C-Flip, FasRBevacizumab, β-catenin, VEGF-A, Slug, NFКB, C-Flip, FasR
Chemical control unit, National Organization for Research and Control of Biologicals, Cairo, Molecular Biology unit, Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Lot release unit, National Organization for Research and Control of Biologicals, Cairo, Department of Cancer Biology, National Institute of Cancer, Cairo University, Department of Applied Research, Research & Development Sector, VACSERA, Cairo, Department of Cancer Biology, National Institute of Cancer, Cairo University, Department of Zoology, Faculty of Science, Cairo University