In vitro experiments, CCK8, colony formation and flow cytometry assays were used to evaluate the anti-
ovarian cancer effect of
damnacanthal on SKVO3 and A2780 cells. The wound healing tests and the transwell invasion assays were used to detect the migration and infiltration of
ovarian cancer cells. Western Blot assays and immunofluorescence staining were used to measure autophagy levels. In vivo experiments, the anti-
ovarian cancer effect of
damnacanthal was further evaluated in a xenograft nude mouse model of SKVO3 cells.
RESULTS:
Damnacanthal induced significant cell death and apoptosis, as well as significant inhibition in migration and invasion, in SKVO3 and A2780 cells, Furthermore,
damnacanthal induced cell cycle arrest by increasing the
protein levels of p27Kip1 and decreasing
cyclin D1 levels. In addition,
damnacanthal induced a significant accumulation of autophagosomes, accompanied with an increase in LC3II
protein levels, and a decrease in p62
protein levels.
3-methyladenine, an autophagy formation inhibitor, significantly mitigated the
damnacanthal-induced apoptosis and migration hindrance, as well as the decline in cell viability. Furthermore, the inactivation of ERK and its downstream effector mTOR signaling pathways, rather than Akt or P38 pathway, were involved in
damnacanthal's activation in autophagy. In addition,
TBHQ, an ERK activator, significantly inhibited
damnacanthal-boosted LC3 II levels and autophagosome accumulation, and reversed
damnacanthal-induced cell death, apoptosis, cell cycle arrest and migration hindrance. Finally, the anti-
ovarian cancer effect of
damnacanthal was confirmed in the orthotopic xenograft model of SKVO3 cells in nude mice, with
tumor growth being significantly inhibited comparably to the efficacy of
cisplatin.
Damnacanthal was also synergistic with
cisplatin and showed inhibition in
cisplatin-resistant
ovarian cancer cells.
CONCLUSION: