The Research On The Reversal Effect Of Psoralen On Glutathione-S-transferase-? Mediated Multidrug Resistance In Breast Cancer

Objective:People around the world work hard to explore the treatment of breast cancer,the main treatment include surgery,chemotherapy,radiotherapy,targeted therapy and so on.Chemotherapy is the most important treatment of breast cancer treatment,preoperative effect is better,but the effect of late chemotherapy is not good.Multidrug resistance of breast cancer has led to thousands of female patients to death,explore and solve the resistance of breast cancer drug-resistant cells to chemotherapy drugs to improve clinical chemotherapy effect,improve the prognosis of patients,improve the survival rate get more and more attention.To explore the molecular basis of multidrug resistance and develop clinical reversal drug to prevent the occurrence of drug resistance is imminent.GST-? plays an important role in enzyme-mediated breast cancer resistance.The reversal effects of Psoralen are clear,whether Psoralen can affect the expression of GST-? has an impact on its function,whether its expression was regulated by other signaling pathways,with this issue we have explored the research.Methods:1.The activity of drug-resistant cell line cells was detected by MTT assay,and the effect of Psoralen on cell proliferation was further detected.2.The expression of GST-? was detected by RT-PCR and agarose gel electrophoresis after treated with different concentrations of Psoralen for 48h.3.Western blot was used to detect the expression of GST-? in MCF-7/ADR cells after treated with different concentrations of Psoralen for 48h.4.Immunofluorescence microscopy was used to observe the activation of NF-?B after Psoralen treatment.5.Western blot was used to detect the expression of NF-?B protein in breast cancer resistant cells MCF-7/ADR after treated with Psoralen for 48h.6.The RT-PCR technology was used to detect the expression of GST-? gene between Psoralen and NF-?B inhibitor treatment groups.7.CCK-8 detection technology was used to detect the difference of adriamycin IC50 between Psoralen and NF-?B inhibitor treatment groups.Result:1.We measured Psoralen can inhibit the proliferation of breast cancer cells byMTT assay,compared with the control group the difference was statistically significant(P<0.05).2.Compared with cells in the control group(without Psoralen treatment group),the GST-? gene was down-regulated in MCF-7/ADR cells treated with different concentrations of Psoralen treatment,difference was statistically significant(P<0.05).There was no significant difference in mRNA expression between different Psoralen groups(P>0.05).The experimental results were consistent with those of agarose gel electrophoresis.NF-?B inhibitor and Psoralen can reduce the expression of GST-? gene.3.The expression of GST-? protein was significantly lower than that of the blank control group(without Psoralen treatment group),but there was no significant difference between the two groups(P>0.05).4.Immunofluorescence microscopy showed that a large amount of NF-?B protein was accumulated around the nucleus after treated with Psoralen for 48 hours.5.After treated with Psoralen at concentration of 8?g/ml,the extraction of nuclear protein of NF-?B expression levels were significantly reduced.6.The expression of GST-? was significantly decreased in MCF-7/ADR cells treated with Psoralen.NF-?B inhibitor and Psoralen can reduce the expression of GST-? gene,NF-?B inhibitor and Psoralen combined treatment group has the strongest effect.7.Psoralen increases the sensitivity of drug-resistant cell lines to chemotherapeutic agents by inhibiting "NF-?B activation.Conclusion:1.The results of MTT assay showed that Psoralen had the effect of inhibiting the proliferation of MCF-7/ADR cells,and the inhibitory effect was dose-dependent.2.The expression of GST-? gene was significantly decreased after treated with different concentrations of Psoralen in MCF-7/ADR cell lines,but there was no dose-dependency on Psoralen concentration.3.Different concentrations of Psoralen treated MCF-7/ADR-resistant cell lines significantly reduced the expression of GST-?,but there was no dose-dependency on Psoralen concentration.4.Psoralen can inhibit NF-?B activation,prevent its transfer to the nucleus,inhibit its function,can reverse drug resistance.5.Psoralen can significantly inhibit the activation of NF-?B into the nucleus,can be used as NF-?B inhibitor.6.Different concentrations of Psoralen significantly reduced GST-? expression levels,but there were no dose-dependent for Psoralen concentrations.Specific NF-?B inhibitors also significantly reduced the expression level of the drug-resistant gene GST-?.Both drugs have the effect of reversing drug resistance and improving the effect of chemotherapy.7.NF-?B specific inhibitor can increase the sensitivity of breast cancer drug-resistant cell lines to chemotherapy drugs,Psoralen also have this effect,GST-? may be a key factor of the NF-?B signaling pathway downstream.