The current study was undertaken to examine the anticancer potential of davanone against human ovarian cancer cells along with evaluating its effects on cell apoptosis, PI3K/AKT/MAPK signaling pathway and cell migration and invasion.
CCK-8 assay was performed for cell viability and clonogenic potential was examined through clonogenic assay. Acridine orange (AO)/Ethidium bromide (EB) dual staining assay was performed to detect apoptosis and quantification of apoptosis was achieved through annexin V-FITC/propidium iodide (PI) staining assay. Mitochondrial membrane potential (MMP) was studied via flow cytometric analysis of ovarian cancer cells. Cell migration and invasion potential of ovarian cancer cells was monitored via transwell assay. Western blotting technique was used to study PI3K/AKT/MAPK pathway.
The results indicated that davanone induced dose as well as time dependent inhibition in cell viability of OVACAR-3 cells. Next, AO/EB staining suggested that the antiproliferative effects of davanone are apoptosis-mediated. There was a remarkable increase in apoptotic cell percentage with the molecule dose. Caspase-3, -8 and -9 activity along with Bax activity were observed to be increasing with davanone doses and Bcl-2 activity decreased with increasing molecule concentration. Transwell assay indicated potential inhibition of invasive and migratory ability of OVACAR-3 cells after davanone exposure. Finally, western blotting analysis revealed that davanone resulted in blocking of PI3K/AKT/MAPK signaling pathway in OVACAR-3 cells.
The results indicate that davanone is a potential anticancer agent against human ovarian cancer mediated via caspase-dependent apoptosis, loss of MMP, inhibition of cell migration and invasion and targeting PI3K/AKT/MAPK signaling pathway.