Abstract
Despite the tremendous efforts to implement new paradigms for breast cancer, the disease still remains a major challenge worldwide. Genetic deregulation is evident in all breast cancer subtypes and comprises a multitude of mutated genes and deregulated signaling cascades. In this sense, co-targeting Src and COX-2 signaling cascades have attracted fervent interest. This work explored the probable anti-carcinogenic effects of Dasatinib as a Src inhibitor, Celecoxib as a selective COX-2 inhibitor, and their combination in MDA-MB-231 triple-negative breast cancer cell line. Drug growth inhibition 50 (GI50) was determined using the MTT assay and the obtained results were analyzed using CompuSyn 3.0.1 software. MDA-MB-231 cells were divided into four treatment groups including a positive control, Dasatinib-treated, Celecoxib-treated, and combination-treated groups. Standard sandwich ELISA was used for the determination of the protein levels of c-Src, Bcl-2, p-AKT, FAK, PGE2, VEGF, and cyclin D1. Active caspase-3 was determined colorimetrically and the expression of COX-2 and c-Src genes was quantitatively determined via quantitative real-time polymerase chain reaction. The GI50 for Dasatinib was 0.05699 µM while that for Celecoxib was 69.0976 µM. Dasatinib up-regulated c-Src gene while Celecoxib and Dasatinib/Celecoxib combination down-regulated such expression level. COX-2 gene was down-regulated by Celecoxib while it was up-regulated by both Dasatinib and Dasatinib/Celecoxib combination. On one hand, Dasatinib, Celecoxib, and their combination significantly reduced the protein levels of c-Src, Bcl-2, p-AKT, FAK, PGE2, VEGF, and cyclin D1. On the other hand, they elevated active caspase-3. To sum up, Dasatinib/Celecoxib combination increased the capability for apoptosis and suppressed proliferation, angiogenesis, migration, and invasion suggesting a strong cross-talk between Src signaling cascade and COX-2/PGE2 via the intermediate PI3K/AKT/mTOR pathway. Further in-vitro and in-vivo studies are warranted to verify the present findings.
Research on the Mechanism of Pulsatilla Potentially Useful for the Treatment of Triple Negative Breast Cancer Based on Network Pharmacology
