Erchen Decoction Interferes With The Multidrug Resistance Of Non-small Cell Lung Cancer Through JNK Signaling Pathway

Background:Lung cancer is the leading cause of morbidity and mortality in malignant tumors,of which non-small cell lung cancer(NSCLC)accounts for 80%of all lung cancers.Its characteristics are short survival and insidious course of disease.Surgery,chemotherapy and radiotherapy,targeted therapy and immune therapy are the main methods of Western medicine in treating NSCLC.Chemotherapy is the cornerstone of the treatment of advanced lung cancer.One of the main reasons for the failure of chemotherapy for lung cancer is the generation of multi-drug resistance(MDR)in lung cancer.Numerous studies have shown that the mechanism of multidrug resistance is closely related to the expression of P-gp and MRP1 proteins.Previous experiments showed that Erchentang could inhibit the activity of JNK and p38 pathways,and the expression of P-gp and MRP protein was closely related to JNK and p38 signaling pathways.However,whether Erchen Decoction can participate in the expression of P-gp and MRP proteins through JNK signaling pathway to reverse multi-drug resistance.In this study,the proliferation of cisplatin-resistant human lung cell line A549/DDP and the expression of P-gp and MRP1 proteins were investigated by in vitro experiments.It will further provide reasonable and accurate theoretical basis for Erchentang in clinical treatment of lung cancer.Objective:To study the effect of Erchentang on A549/DDP cell viability,apoptosis and related multi-drug resistance proteins P-gp and MRP1 in vitro,so as to provide theoretical basis for Erchentang in the treatment of lung cancer.Research methods:1.MTT method was used to detect the growth inhibition of Erchentang and Erchen tang+Liuwei Formula,Erchentang Simplified Formula,Erchentang+Yiqi Yangyin Formula on A549/DDP cells 48 hours later,so as to screen out the better drug concentration as the basis for the next experiment.2.The immune fluorescence technique was used to observe the effects of DDP group,Erchentang group and Erchentang+DDP group on the morphology of A549/DDP cell line and the expression of P-gp protein.3.Western Blot(WB)was used to detect the effects of DDP group and Erchentang group,Erchentang+ DDP group on the morphology of A549/DDP cell line and the expression of P-gp and MRP1 protein.Results:1.MTT results showed that different Erchentang with 25.6 mg/ml concentration,inhibited A549/DDP cell lines,and compared with the blank control group,the difference was statistically significant(P<0.05).The Erchentang group(25.6mg/ml)had significant difference compared with the blank control group(P<0.01).Erchentang(25.6 mg/m)had significant difference compared with cisplatin(lOug/ml)group(P<0.05).Therefore,the Erchentang group with 25.6mg/ml concentration was selected to complete the further experiment.2.Immunofluorescence technique showed that the morphology of A549/DDP cells in blank control group and DDP group did not change,and the distribution of P-gp protein in cytoplasm was relatively close.In A549/DDP cells containing Erchentang group,nuclear morphology atrophy was observed,P-gp protein expression in cytoplasm was weak,and fluorescence was evacuated.The morphological changes of A549/DDP cell lines were more obvious in Erchentang+DDP group.3.Western Blot test results showed that compared with the blank control group,the expression of multidrug resistance-related proteins P-gp and MRP1 in Erchentang group were significantly lower(P<0.05).In P-gp protein expression:Erchentang+DDP group compared with the blank control group had significant difference(P<0.01),compared with DDP group,the difference was also statistically significant(P<0.05).There was significant difference between Erchentang group and blank control group in detecting MRP1 protein expression(P<0.05).Conclusion:1.Different Erchentang prescriptions can inhibit the activity of A549/DDP cells when the concentration of Erchentang is 25.6 mg/ml.2.Erchentang can reverse multi-drug resistance in cancer cells,and its mechanism is related to the decrease of the expression of multi-drug resistance-related proteins P-gp and MRP1 through JNK signal pathway.