435eP Targeting PGK1 sensitizes urothelial carcinoma to immunotherapy

Background: Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer, with the highest mortality rate and no effective treatments currently available.Research into new therapies, including dietary approaches, is urgently needed.Epidemiological studies suggest that consuming Brassicaceae vegetables, rich in isothiocyanates, may help reduce cancer risk.In our recent study, we demonstrated that Erucin (ERU), the most abundant hydrogen sulfide (H 2 S)-releasing isothiocyanate found in arugula (Eruca sativa), exerts a concentration-dependent inhibition of MDA-MB-231 TNBC cell proliferation by inducing apoptosis and autophagy.ERU also reduced ROS levels, enhanced antioxidant gene expression, and blocked cell migration, invasion, and colony formation.In the present study, we explored the potential anti-cancer effects of ERU in vivo.Methods: Female BALB/c mice were injected with 4T1 clone 5 cells to assess tumor growth, spleen weight, and metastatic burden.ERU (10 mg/kg) was administered orally once daily for 28 days, after which the mice were sacrificed.Tumors were collected and analyzed using RT-PCR, western blotting, and flow cytometry. Results:Our results revealed ERU significantly reduced tumour volume and weight in 4T1 mice.Moreover, ERU affected the NF-B signalling pathway, reducing the expression of several inflammatory mediators and enzymes, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumour necrosis factor-alpha (TNF) and interleukin-6 (IL-6).Furthermore, ERU inhibited angiogenesis in 4T1 tumours as revealed by the reduced expression of vascular endothelial growth factor (VEGF).Preliminary immunophenotyping analysis showed that ERU reduced the number of monocytes (CD45 + Cd11b + Ly6G -Ly6C + ) and CD8 + PD1 + T cells within the tumor. Conclusions:In conclusion, our results show that the consumption of ERU could be a useful dietary support intervention to prevent and treat TNBC.