Radiosensitization And Pharmacokinetics Of Wogonin On Breast Cancer

Scutellaria baicalensis?Huang Qin?as an effective medicine active in Huang qin Tang.It is reported to be effective on breast cancer and it's active ingredient,wogonin,also has anti-cancer effects in breast cancer.The previous work of our group found that wogonin can effectively inhibit the Nuclear Factor E2-related factor 2?Nrf2?signaling pathway.It has been reported that Nrf2 may be a potential target for radiosensitization,which indicates that wogonin may have a potential sensitization effect on breast cancer.In this paper,intends to use breast cancer MCF-7 and MDA-MB-231 cells to establish a radioresistance model to study whether wogonin has radiosensitization effect on breast cancer.And established a High performance liquid chromatography-tandem mass spectrometry?HPLC-MS/MS?method to simultaneously determine wogonin and docetaxel in rat plasma.Then,this method was used to explore the pharmacokinetics of wogonin and docetaxel in rats with mammary tumors.methods:1.Radioresistance MCF-7R and MDA-MB-231R were derived after fractionated irradiation of parental cells with a total dose of 60 Gy?2 Gy per fraction,five times per week?.Using colony formation assay multi-target single-hit model to verify the success of the radiotherapy resistance model.Tetrazolium salt Colorimetric?MTT?Method was used to measure the inhibitory effect of wogonin on the proliferation of MCF-7R and MDA-MB-231R cells.Using colony formation assay to detection whether wogonin had radiosensitization effect on MCF-7R and MDA-MB-231R cells.2.Establishing a HPLC-MS/MS method to simultaneously determine wogonin and docetaxel concentrations in rat plasma,and the Methods Validation was examined.3.Using N-nitroso-N-methylurea?NMU?to induce mammary tumors in rats.After the administration of wogonin and docetacxel in rats with mammary tumor,The plasma concentrations were determined,and the pharmacokinetic characteristics of wogonnin and docetaxel in rats with mammary tumors were studied.Results:1.Radiosensitivity parameters SF₂?MCF-7?=60.64%and SF₂?MCF-7R?=81.74%?P<0.01?;SF₂?MDA-MB-231?=26.82%and SF₂?MDA-MB-231R?=42.58%?P<0.01?.The IC50 values of wogonin on MCF-7R and MDA-MB-231R cells for 48h were104.74±2.4?M and 141.40±2.8?M,respectively.Wogonin at concentrations of 25?M and 50?M both have a radiosensitization effect on MCF-7R and MDA-MB-231R cells.2.The established HPLC-MS/MS method uses paclitaxel as an internal standard,The chromatographic separation was accomplished on a HypersilGold?C₁₈ column?5?m,2.1×150 mm i.d,Agilent,USA?and the column temperature was set at 30°C,acetonitrile and 0.1%formic acid in water?60:40,v/v?as mobile phase,The flow rate was set at 0.3 mL/min.For mass spectrometer,positive ionization is adopted.The results of Methods Validation showed that the linear range has good correlation?R²>0.99?,The intra-and inter-day precision?%RSD?with in 15%,Accuracy is 85%-115%,the recoveries were 72.38%-83.25%and the matrix effects were 92.83%-105.23%,The sample has good stability.3.The rat with mammary tumors was successfully established after 35 days,by intraperitoneal injection of NMU 75mg/kg.The C_max and AUC₀–_t of wogonin were proportionally increased in the dose range from 10 to 40 mg/kg.Moreover,the C_max?AUC₀–_t and t_1/2/2 of docetaxel and the AUC₀–_t of wogonin were increased after co-administration?p<0.05?.Conclusion:The radioresistance models of MCF-7R and MDA-MB-231R cells were established successfully by fractionated irradiation of parental cells with a total dose of 60 Gy?2 Gy per fraction,five times per week?.wogonin can radiosensitization MCF-7R and MDA-MB-231R cells.The established HPLC-MS/MS method for simultaneously determine wogonin and docetaxel concentrations in rat plasma was rapid,accurate,stable,and specific,which could be used for the subsequent study on the pharmacokinetic effects of wogonin on docetaxel in rats.Pharmacokinetic interactions between wogonin and docetaxel exist in vivo when used together,which could result in a significant increase in both of their in vivo exposures.Therefore,there is a need to consider the change in pharmacokinetics when both of them are used together.