| Background and objectives: Epithelial Ovarian cancer(EOC,Ovarian cancer)is one of the three most common malignant tumors in gynecology.is the most common histological type of ovarian malignant tumors.Studies have shown that MyD88 positive in about 77.1% of ovarian cancer cells,and it’s an independent prognostic factor of epithelial ovarian cancer.In epithelial ovarian cancer cells,the formation of TLR4/MD-2 complex can activate MyD88,thereby increasing the secretion of pro-inflammatory cytokines(IL-6,VEGF,IL-17 A,etc.)and related to paclitaxel resistance,affecting the tumor-free survival period and overall survival of patients with epithelial ovarian cancer.Pharmacological studies have shown that Sulforaphane(SFN)has anti-tumor effects.SFN has been found to inhibit the formation of TLR4/MD-2 complex on the cell membrane.Whether SFN can inhibit MyD88 positive ovarian cancer cells and its specific mechanism are still poorly reported.we used MyD88 negative ovarian cancer cell A2780 and MyD88 positive ovarian cancer cell OVCAR-3 to study the specific mechanism of SFN on MyD88 positive ovarian cancer cells,to provide a direction of study for the treatment of ovarian cancer and chemosensitizing drugs,and improve the quality of life and overall survival rate of these patients.Methods:1.Flow cytometry was used to detect the formation of TLR4/MD-2 complex on the membrane of OVCAR-3 ovarian cancer cells,which induced by SFN,LPS,and SFN combined with LPS for 6 hours.2.RT-q PCR was used to detect the expression of MyD88 mRNA and IL-6 mRNA in OVCAR-3 cells and A2780 cells after induced by LPS,PTX,SFN and PTX combined with SFN respectively for 6 and 12 hours.Results:1 、 After 6 hours of LPS co-culture with OVCAR-3 cells,the expression ofTLR4/MD-2 was higher than that in the control group by flow cytometry.2、After co-culture of SFN and OVCAR-3 cells at different concentrations for 6 hours,the TLR4/MD-2 complex was significantly reduced compared with the control group by flow cytometry,and in a concentration-dependent manner.3、The TLR4/MD-2 complex was significantly reduced after the OVCAR-3 cocultured with LPS combination with SFN for 6 hours.4、MyD88 mRNA and IL-6 mRNA was expression both in ovarian cancer cells A2780 and OVCAR-3.Compared with the cell’s of OVCAR-3,the level of MyD88 mRNA and IL-6 mRNA in A2780 cells was very low.After 6 and 12 hours of treatment with LPS、SFN、PTX and PTX combined with SFN,compared with the control group,the level of MyD88 mRNA and IL-6 mRNA expression in A2780 cells had no significant change.5、After culture with LPS or PTX at different concentrations for 6 and 12 hours,the expression of IL-6 mRNA in OVCAR-3 cells was significantly higher than that in the control group,and in a time-dependent manner;but A2780 cells not have the similar change.6、After culture with SFN in different concentrations for 6 and 12 hours,the expression of IL-6 mRNA and MyD88 mRNA in OVCAR-3 cells decreased significantly compared with that in control group,and in a time and concentrationdependent manner.7、After culture with SFN combined with PTX for 6 and 12 hours,the expression of MyD88 mRNA IL-6 mRNA in OVCAR-3 cells decreased significantly compared with that in PTX group.Conclusions:SFN can inhibit the formation of TLR4/MD-2 complex on the membrane of MyD88 positive ovarian cancer cells and decrease the expression of MyD88 mRNA and IL-6 mRNA in this cells;inhibiting the formation of TLR4/MD-2 complex on the membrane of cells induced by LPS;antagonize the increase of MyD88 mRNA and IL-6 mRNA expression in cells induced by PTX.Therefore,SFN can inhibitTLR4/MyD88 signaling pathway and the expression of downstream cytokine IL-6 mRNA to inhibit the proliferation of ovarian cancer and sensitize paclitaxel. |
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