Sulforaphrane Down-regulates Paclitaxel-mediated Expression Of PD-L1,IDO1 And AHR And Activity Of IDO1 In MyD88~+ Human Ovarian Cancer Cells

Background and objectives:Epithelial ovarian cancer(EOC)is the highest mortality female genital tract malignant tumors worldwide,accounting for 85% to 90% of ovarian malignant tumors.More than 60% of the patients were diagnosed as advanced tumor due to the vague early symptoms and the lack of screening methods.Although new progress has been made in the treatment of ovarian cancer,the five-year survival rate of patients with advanced ovarian cancer is still about 30%.Studies have confirmed that immunotherapy is a promising therapeutic strategy in basic research and animal research in recent years,and EOC as an immunogenic tumor may benefit from it.It has been found that programmed death receptor ligand-1(PD-L1),indoleamine2,3-dioxygenase1(IDO1)and aromatic hydrocarbon receptor(AHR)are highly expressed in many malignant tumors,including ovarian cancer,and are associated with immune escape and poor prognosis.Therefore,inhibiting the expression and activity of PD-L1,IDO1 and AHR may improve the immunosuppression and prognosis of ovarian cancer.AS a natural isothiocyanate compound in the seeds of raphani,sulforaphrane(SFN)has been shown that to inhibit various carcinogenic signaling pathways in human cancer,including nuclear factor-kappa B(NF-kappa B),Akt and various survival pathway proteins.However,whether it can down-regulate the expression of PD-L1,IDO1 and its downstream factor aromatic hydrocarbon receptor through My D88/NF-kappa B pathway remains to be explored.In this study,the effects of SFN on the expression of PD-L1,IDO1,AHR and the activity of IDO1 in human ovarian cancer cells were studied.Methods:1.Real-time fluorescence quantitative PCR(rt-pcr)was used to detect the expressions of pd-l1,IDO1 and AHR m RNA in My D88+ OVCAR-3 cells and My D88-A2780 cells at different concentrations of SFN at different time points;2.Colorimetry was used to detect the changes of kyn concentration in supernatants of My D88+ OVCAR-3 and My D88-A2780 cells before and after the effecst of SFN.Results:1.Both OVCAR-3 and A2780 cells expressed PD-L1,IDO1 and AHR,and the expression in OVCAR-3 cells was significantly higher than that in A2780 cells.2.SFN down-regulated the expression of PD-L1 in OVCAR-3 cells in a concentration-dependent manner,with the most obvious down-regulation after 6hours of 10 umol/L treatment,and reduced the expression of PD-L1 induced by PTX.3.SFN down-regulated the expression of IDO1 in OVCAR-3 cells in a time-dependent manner,especially after 12 hours,and reduced the expression of IDO1 induced by PTX.4.SFN down-regulated the expression of AHR in OVCAR-3 cells in a time-dependent manner,especially after 12 hours,and reduced the expression of AHR induced by PTX.5.SFN reduced Kyn secretion in OVCAR-3 and A2780 cells.Conclusions:1.The overexpression of PD-L1,IDO1 and AHR in OVCAR-3 cells may be related to immune escape of ovarian cancer.2.SFN inhibits the transcription of PD-L1,IDO1 and Ah R,but the optimal time and concentration of them are different.SFN combined with PTX can inhibit the expression of PD-L1,IDO1 and AHR induced by PTX,and enhance the response of My D88+EOC cells to PTX.3.SFN can reduce the secretion of kyn in OVCAR-3 cells,and the secretion after combined with 1-methyl tryptophan(1-MT)is significantly lower than that of SFN alone.It shows that the use of 1-MT and TLR-4/My D88 upstream inhibitors can synergistically inhibit IDO1,which may have a positive clinical significance.4.SFN can improve the immunosuppressive status of ovarian cancer,especially OVCAR-3 cells.Inhibiting the immune escape of ovarian cancer may improve the prognosis of ovarian cancer patients.