To Investigate The Clinical Efficacy And Mechanism Of Qiyu Sanlong Prescription Combined With Chemotherapy In Patients With Advanced NSCLC

Objective1.To observe the effect of Qiyu Sanlong prescription on the clinical efficacy and quality of life of patients with advanced non-small cell lung cancer(NSCLC)by retrospective study.2.To investigate the mechanism of Qiyu Sanlong prescription on the proliferation,invasion and migration of lung cancer cells.Methods1 Clinical TrialA total of 70 patients with advanced NSCLC who met the inclusion criteria were selected from Anhui Provincial Hospital of Traditional Chinese Medicine and the First Affiliated Hospital of Anhui Medical University from September 2020 to September2022.According to the clinical history,the patients were divided into two groups,35cases in chemotherapy alone group and 35 cases in combined treatment group.According to the clinical history,the patients were divided into two groups,35 cases in chemotherapy alone group and 35 cases in combined treatment group.Pure chemotherapy group with chemotherapy alone,combined treatment group with chemotherapy combined Qiyu Sanlong prescription treatment.Describe the two groups before treatment in patients with solid tumor size,tumor markers,TCM syndrome score,scores and adverse reaction of functional status,and comparing with the corresponding changes after treatment.2 Experimental ResearchThe lyophilized powder of Qiyu Sanlong prescription was prepared according to the previous research.Mouse LLC cells were cultured in vitro.The LLC cell line stably overexpressing Wnt5a was constructed by lentivirus transfection,and the construction and culture of Wnt5a interference and empty vector cell lines were completed at the same time.The expressions of Wnt5a andβ-catenin were detected by q RT-PCR.With CCK8 and EDU technology to detect cell activity,breeding and genetic ability.Transwell assay was used to detect cell migration and invasion.The protein expressions of Wnt5a,β-catenin,c-Myc,Cyclin D1,Vimentin,N-cadherin,E-cadherin and Snail were analyzed by Western blotting.Results1 Clinical TrialIn this study,70 patients were enrolled,3 were excluded and 2 were shed,and a total of 65 patients were finally included in the study.In all patients,included in the combined treatment group 32 cases,33 cases of chemotherapy alone group.(1)General data include gender,age,BMI,cancer pathological classification and TNM stage.Statistical analysis of them shows P>0.05,which can be compared between the two groups.(2)The size of solid tumor was recorded before and after treatment,and the analysis between groups after treatment showed thatX~2=1.382,P=0.501>0.05,and there was no significant difference between the two groups.(3)The levels of SCC,CEA,CA125 and CYFRA21-1 in the two groups before and after treatment were recorded.After analysis,the indexes in the two groups except CYFRA21-1 decreased to varying degrees(P<0.05).The level of CYFRA21-1 in the control group after treatment was lower than that before treatment,and there was no significant difference in the treatment group.(4)The TCM syndrome score was comprehensively evaluated from the aspects of cough,chest pain,fatigue and so on.Before treatment,the scores of the above aspects were statistically analyzed between the treatment group and the control group,P>0.05,there was no statistically significant difference between the two groups.After treatment,cough,sputum blood,chest tightness and shortness of breath were improved in both groups,and the improvement of cough in the treatment group was better than that in the control group.The treatment group was better than the control group in chest pain,fatigue,loss of appetite,spontaneous sweats and night sweats,dry throat and dry mouth.(5)In terms of functional status,we need to perform KPS scores on the treatment group and the control group before treatment to clarify whether there are functional differences between the two groups before treatment.Statistical results show P=0.748>0.05.The two groups can compare the data after treatment.After treatment,compared with the control group,P=0.046<0.05 before and after treatment in the treatment group,indicating that Qiyu Sanlong prescription combined with chemotherapy could reduce patients KPS score,improve the function of patients.(6)There was no significant difference in bone marrow suppression,liver and kidney function between the two groups.In terms of gastrointestinal reaction,the comparison between the two groups was P<0.05,and the combined treatment group could significantly improve the symptoms of nausea and vomiting.During the treatment,there were no nervous system,heart,infection,allergy,skin and other toxic reactions in the two groups.2 Experimental Research(1)Cell proliferation ability:CCK8 test revealed that Qiyu Sanlong prescription was concentration-dependent under different concentrations and different time comparisons.With the increase of drug concentration,the inhibition rate of Qiyu Sanlong prescription on lung cancer cells increased continuously.Ed U experiment was used to detect DNA synthesis of genetic material in cells after drug treatment.Imaging analysis showed that compared with the control group,the positive rate of Ed U in cells of QYSL group decreased significantly,and cell proliferation was inhibited.(2)Cell migration and invasion ability:In Transwell assay,lung cancer cells were seeded and cultured in different nutrient media for 24 hours.Microscopically,the cells in the lower chamber were counted and photographed.It was found that the number of cells in the QYSL group was significantly lower than that in the control group,with statistical significance between group comparisons and before and after treatment.(3)Construction of Wnt5a overexpression and interference model:overexpressed lentivirus and si RNA were used to transfuse Wnt5a in lung cancer cells.To confirm the success of the model construction,the expression levels of Wnt5a andβ-catenin in lung cancer cells were detected by q RT-PCR.Wnt5a overexpression and interference cell lines were successfully and stably constructed.(4)Protein expression levels of Wnt/β-catenin pathway:Western blot was used to detect the protein levels of the pathway in lung cancer cells.The protein expressions of Wnt5a,β-catenin,Cyclin D1 and C-Myc in the QYSL group were lower than those in the oe-Wnt5a group.After transfection,the expression of Wnt5a and other related proteins in the oe-Wnt5a group increased,and the protein expression decreased after the drug was added.The opposite was observed in the si-Wnt5a group.(5)Protein expression level of EMT process:Protein expression was measured in the oe-Wnt5a group and the si-Wnt5a group respectively.The expression of N-cadherin,Vimentin and Snail increased and the expression of E-cadherin decreased in the oe-Wnt5a group,while the opposite was observed in the si-Wnt5a group.After drug treatment,the expression of related proteins in the oe-Wnt5a group was reversed.Conclusion1 Qiyu Sanlong prescription combined with chemotherapy can effectively improve the TCM syndromes and physical status of patients with stageⅢ-ⅣNSCLC,reduce the expression of some tumor markers and the toxic side effects of chemotherapy.The addition and modification of Qiyu Sanlong decoction with syndrome is of great significance in improving the quality of life,chemotherapy tolerance and compliance of patients.2 Qiyu Sanlong prescription can inhibit the expression of Wnt5a/β-catenin signaling pathway related proteins,reduce the translocation ofβ-catenin protein,increase the expression of N-cadherin protein in mesenchymal cells,decrease the expression of E-cadherin protein in epithelial cells,and lose the adhesion junction between cells,thereby inhibiting the initiation of EMT in lung cancer cells.Inhibition of lung cancer cell metastasis.