Combining Bevacizumab with Knocked-Down β-Catenin Reduces VEGF-A and Slug mRNA in HepG2 but not in Caco-2 Cell Lines
Reem Mebed1, Yasser Bm Ali2, Nahla Shehata1
1Egyptian Drug Authority, CA of Biological and Innovative Products and clinical studies, Cairo, Egypt.
Background:
Bevacizumab (Bev) resistance is hypothesized to be overcome by combining 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, NFκB, 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, 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 6) 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 treated HepG2 cells. In contrast, VEGF-A and Slug mRNA levels in Bev 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-catsiRNA remarkably reduced the level of NFκB, FasR, and c-Flip compared to Bev treated HepG2 cells, although the difference was not statistically significant.
Conclusion:
We conclude that combining Bevacizumab with knocked down β-catenin reduces the expression of VEGF-A and Slug in HepG2 but not in Caco-2 cells.
Insights
Combining Bevacizumab with beta-catenin knockdown reduced VEGF-A and Slug expression in HepG2 cells. This combination therapy shows potential for overcoming Bevacizumab resistance by targeting key signaling pathways.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Bevacizumab (Bev) resistance is a significant challenge in cancer therapy.
- Combining inhibitors of other signaling pathways is a potential strategy to overcome Bev resistance.
Purpose of the Study:
- To investigate the effect of combining Bevacizumab with beta-catenin knockdown on key gene expression in cancer cells.
- To assess the impact on VEGF-A, Slug, NFκB, c-Flip, and FasR expression in HepG2 and Caco-2 cell lines.
Main Methods:
- Cultured HepG2 and Caco-2 cells were treated with Bevacizumab, beta-catenin siRNA, or a combination (Bev-β-cat-siRNA).
- Gene and protein expression levels were analyzed using quantitative PCR (qPCR) and western blotting.
- Control groups included untreated cells and cells treated with negative controls.
Main Results:
- Bev-β-cat-siRNA significantly reduced VEGF-A and Slug mRNA in HepG2 cells, contrasting with Bev treatment alone.
- VEGF-A and Slug expression patterns differed between HepG2 and Caco-2 cells.
- A notable reduction in NFκB, FasR, and c-Flip levels was observed with Bev-β-cat-siRNA in HepG2 cells.
Conclusions:
- Combining Bevacizumab with beta-catenin knockdown effectively reduces VEGF-A and Slug expression in HepG2 cells.
- The observed effects were cell-type specific, with different outcomes in Caco-2 cells.
- This combination strategy may offer a novel approach to combat Bevacizumab resistance.
