The Clinical Trial And Experimental Mechanism Of Fuzheng Kangai Decoction And Gefitinib On Non-small Cell Lung Cancer

Objective:Clinical Research:To evaluate the efficacy of Fuzheng Kangai Decoction and Gefitinib in the treatment of advanced non-small cell lung cancer(NSCLC)patients with EGFR gene mutation and to alleviate its toxicity and side effects.Experimental research:To explore the molecular mechanism of Fuzheng Kangai Decoction and Gefitinib in inhibiting the proliferation of lung cancer cells through DNMT1/SP1/MET pathway.Method:Clinical Research:Seventy advanced non-small cell lung cancer(NSCLC)patients with EGFR gene mutation were randomly divided into experimental group(Gefitinib+Fuzheng Kangai Granule)and control group(Gefitinib+placebo).Treatment were given until cancer progression,intolerable toxic side effects or death.To observe the progression-free survival(PFS),disease control rate(DCR),quality of life(EORTCQLQ-LC43),overall survival(OS)and toxicity of two groups of patients,in order to understand whether Fuzheng Kangai Decoction and gefitinib can improve the efficacy of Gefitinib and reduce its toxicity and side effects in advanced non-small cell lung cancer with EGFR gene mutation.Experimental research:The survival rates of A549 and H1975 cells were detected by MTT assay.The expression of DNA methyltransferase 1(DNMT1),tyrosine protein kinase MET and transcription factor SP1 were detected by Western Blot.The mRNA expression of MET was detected by qRT-PCR,and the exogenous overexpression of DNMT1,SP1 and MET was detected by transient transfection.MET promoter activity was detected by double luciferase reporter gene assay.The experimental results in vitro were validated by nude mice model that transplanted tumors.Results:Clinical Research:The disease control rate(DCR)of the treatment group and the control group were 93.90%and 90.30%respectively.The treatment group was higher than the control group,but there was no significant difference between two groups(P>0.05).The median PFS was 13.5 months in the treatment group and 11.7 months in the control group.There were no significant differences between the two groups(P>0.05).In the stratified analysis of the treatment group,there was no significant difference in gender,age,pathological type,EGFR mutation,TNM stage,PS score and treatment choice in PFS,P>0.05.In terms of smoking,the median PFS of smoking patients and non-smoking patients in the treatment group was 1.8 and 14.6 months,respectively.There were significant differences between the two groups(P<0.05).In the stratified analysis of the control group,there were no significant differences in gender,age,smoking,pathological type,EGFR mutation,TNM stage,PS score and treatment choice in PFS,P>0.05.In terms of quality of life EORTC QLQ-LC43,the treatment group(Gefitinib+Fuzheng Kangai Granule)improved on physical function,role function,emotional function,cognitive function,social function,fatigue,nausea and vomiting,pain,shortness of breath,appetite,constipation or diarrhea(P<0.05).In the control group(Gefitinib+placebo),physical function,role function,emotional function,cognitive function,social function,fatigue,nausea and vomiting,loss of appetite were improved after treatment(P<0.05).In terms of role function,pain and diarrhea,the treatment group ’ patients felt better than the control group after the trial s treatment,with statistical differences(P<0.05).In terms of toxicity and side effects,10 patients(30.3%)in the treatment group and 14 patients(45.2%)in the control group had adverse reactions,all of which were Ⅰ～Ⅲ degree adverse reactions.There were no significant differences between the two groups(P>0.05).The incidence of diarrhea,oral mucositis and abnormal liver-function was similar in the two groups,but the incidence of skin rash in the treatment group was 2 cases,while in the control group there was 8 cases,the incidence of skin rash in the treatment group was lower than the symptom in the control group(P<0.05).Experimental research:We demonstrated that Fuzheng Kangai Decoction and Gefitinib can inhibit the proliferation of human lung cancer cells A549 and H1975.In addition,we found that Fuzheng Kangai Decoction inhibited the expression of DNMT1 and SP1 proteins.Moreover,Fuzheng Kangai Decoction can reduce the expression of protein and mRNA and the activity of MET promoter.Exogenous overexpression of DNMT1 and SP1 can reverse the inhibitory effect of Fuzheng Kangai Decoction on DNMT1,SP1 and MET.Exogenous overexpression of MET can reverse the inhibitory effect of Fuzheng Kangai Decoction on the proliferation of lung cancer cells.In vivo experiments,it also showed that Fuzheng Kangai Decoction inhibited the proliferation of lung cancer cells and decreased the expression of DNMT1,SP1 and MET proteins.Conclusions:1.Fuzheng Kangai Decoction combined with Gefitinib can prolong the progression-free survival of NSCLC patients with EGFR gene mutation,alleviate toxicity and side effects,and improve the quality of life.2.Fuzheng Kangai Decoction and Gefitinib inhibited the growth of lung cancer cells through DNMT1/SP1/MET pathway.At the same time,the expression of DNMT1,SP1 and MET was decreased by Fuzheng Kangai Decoction.With Fuzheng Kangai Decoction,DNMT1 and SP1 can regulate each other and ultimately inhibit MET gene expression.This study partly reveals the new mechanism of Fuzheng Kangai Decoction in inhibiting the growth of lung cancer cells.It also indicated that MET could be used as a target in the treatment of lung cancer.