Introduction/Background Ovarian cancer is one of the three major gynecological malignancies, with the highest mortality rate. The field of cuproptosis research has experienced exponential growth in the past year. While numerous studies have established the association between cuproptosis and various cancers, its relevance to ovarian cancer remains relatively understudied. Our objective is to investigate the potential therapeutic implications of cuproptosis driven by cyclin-dependent kinase inhibitor 2A(CDKN2A)/p16 in ovarian cancer.

Methodology CDKN2A and control siRNAs were transfected into ovarian cancer OVCAR3 and ES-2 cells. qPCR was used to determine the transfection efficiency in mRNA level. 1μM of CuCl2 was added 30 minutes prior to drug administration. To investigate the impact of CDKN2A on cell growth via regulating cuproptosis, cells were pre-treated with 1μM of CuCl2, followed by treatment with either elesclomol (a crucial copper ionophore) alone or elesclomol and copper in a 1:1 ratio. The viability of cells was then assessed by XTT assay. Moreover, given that the oligomerization of lipoylated DLAT is a crucial hallmark of cuproptosis, non-denaturing gel electrophoresis and Western blot analysis were conducted.

Results qPCR results demonstrated a significant reduction in p16 mRNA expression in CDKN2A/p16-si cells when compared to control cells. We also demonstrated that transient knockdown of CDKN2A/p16 resulted in increased sensitivity to elesclomol/elesclomol-Cu treatment in a dose-dependent manner compared to control cells. Similar results were observed in both OVCAR3 and ES-2 cells. Western blot analysis revealed increase oligomerization of lipoylated DLAT accompanied by decrease of DLAT monomers in CDKN2A/p16 si cells after drugs treatment.

Conclusion Reduced CDKN2A/p16 expression sensitized cells to elesclomol/elesclomol-Cu treatment in a dose-dependent manner. CDKN2A/p16 may contribute to cuptoptosis-induced cell death in ovarian cancer.

Disclosures All the authors declare no personal or funding interests.