Curcuminoids WM03 inhibits ovarian cancer cisplatin-resistant cells proliferation and reverses cisplatin resistance by targeting DYRK2

Abstract

Purpose: Cisplatin is a common chemotherapy agent used to treat ovarian cancer and cisplatin resistance is the most common consequence after its treatment. Curcumin has been shown to effectively inhibit the proliferation and invasion of ovarian cancer cells but its bioavailability restricts its application. The objective of this study was to develop the novel curcumin derivatives with high efficacy and synergic effects with cisplatin to inhibit cisplatin resistant ovarian cancers.

Study design and methods: Colony formation assay and growth curve assay Were used to detect cell proliferation. Transwell and cell scratch assay Were used to detect cell invasion and migration. Western blot (WB), Immunohistochemistry (IHC) and Immunofluorescence (IF) Were used to detect the expression levels of related molecules. qPCR was used to detect mRNA levels of related molecules. Kinase profile sequencing was used to analyze kinase activity. RNA seq was used to analyze significant signaling pathways. The ability of Surface plasmon resonance (SPR), Isothermal titration calorimetry (ITC) and Cellular Thermal Shift Assay (CESTA), molecular docking to analyze the binding of drugs and molecules; Co-Immunoprecipitation (Co-IP) and confocal are used to analyze intermolecular interactions. Ubiquitination is used to detect ubiquitin levels of related molecules; Animal experiments are used to simulate clinical validation RESULTS: Four curcumin derivatives Were synthesized and evaluated to treat ovarian cancers. Curcumin derivative WM03 was the most effective to inhibit A2780DR and HO8910PMDR cell proliferation with about 8-12 times more potent than curcumin. WM03 inhibited A2780DR and HO8910PMDR cell proliferation, migration, and invasion with a synergic effect of cisplatin for cisplatin resistant ovarian cells. RNA-seq results showed that the PI3K-Akt pathway differentially changed. Kinotome analysis showed that WM03 specifically targeted 4 kinases of 50 curcumin-effective kinases and dual-specificity tyrosine-(Y)-phosphorylation regulated kinase 2 (DYRK2) was the most significant kinase, The IC₅₀ of WM03 on DYRK2 activity is 4.58 μM, and the strong binding ability of WM03 to DYRK2 was confirmed in cell-free systems such as SPR, ITC and CESTA. Docking analysis showed that WM03 bound to the ATP pocket of DYRK2 similarly to curcumin. Further analysis showed that WM03 significantly inhibited ovarian cell proliferation and invasion via DYRK2-Akt/ATP7A/CTR1 axis. Tumor inoculation in nude mice demonstrated that WM03 at 5 mg/kg every 2 days for 16 days was effective to reduce tumor size.

Conclusion: WM03 specifically targets DYRK2 and is more potent than curcumin to inhibit cisplatin resistant ovarian cancer cells, being a promising new drug candidate for ovarian cancers.

Keywords: ATP7A; CTR1; Curcumin; DYRK2; Ovarian cell proliferation; WM03.