Clinical And Experimental Study Of YQCTF Combined With Concurrent Chemoradiotherapy For Treating Stage ? Lung Squamous Cell Carcinoma

ObjectiveClinical Study objective:To study the effect of YQCTF on ORR and progression-free survival for patients with stage III lung squamous cell carcinoma treated by concurrent chemoradiotherapy,as well as treatment-related side effects,and analyze the efficacy and quality of life of YQCTF on different syndrome type patients,Explore patients with superior efficacy..Experimental research objective:Through animal experiments,we observed the effect of YQCTF on proliferation,apoptosis,cycle and JNK,p-JNK gene of human lung squamous cell carcinoma SK-MES-1 cells,and explored the Possible molecular mechanisms of YQCTF on inhibiting human lung squamous cell carcinoma.method:Clinical study:1 Clinical study methods:A prospective randomized method was used to randomly select 120 patients in the four centers to be treated as the treatment group and 60 in the control group.The treatment group was treated with concurrent chemoradiotherapy+Yiqi Quyu Decoction(in the Qi and Qi On the basis of the prescription,according to the TCM syndromes of lung stagnation,spleen deficiency and dampness,yin deficiency and heat,and qi and yin deficiency,the control group used concurrent radiotherapy and chemotherapy to collect relevant data of patients before and after treatment,including tumor size and blood routine test,liver and kidney function,TCM syndrome type,ECOG PS score,gene(Part of the patients),FACT-L4.0 scale score,etc,after the end of and chemoradiotherapy,review the relevant data to evaluate the efficacy,toxicity and quality of life scale score,every 2 months after chemotherapy,there is a CT scan to assesse tumor changes until tumor progression,recording the time from the onset of the first chemotherapy to the progression of the tumor,defined as PFS.Factors that may affect PFS were selected into the Cox proportional hazard model to screen for factors that have an impact on PFS.2 Statistical methods:Statistical analysis was performed using SPSS software.The measurement data were expressed as mean ± standard deviation(x±S).Paired t test was used for each group before and after each group.For those who did not meet the normal distribution,the rank sum test was used.All count data were frequency(f)and percentage or composition.Ratio(p);two sets of grade data using independent sample rank and Mann-Whitney U test;unordered classification data using Pearson chi-square test(x2),four?40 data but 1?T<5 When using the corrected chi-square test;when n<40 or T<1,use the Fisher exact probability method;Comparison of progression-free survival time(PFS)between groups using Log-rank test;Cox regression analysis,using Backward wald regression analysis for multivariate analysis;all data are counted by SPSS 20.0.The value of a is taken as 0.05.Experimental research method:1 Cell experiments To prepare YQCTF,After resuscitation and passage of human lung squamous cell carcinoma SK-MES-1 cells,the cell count was observed under an inverted microscope.The cell density was calculated by the formula,and the cell viability was detected by CCK8 method.The effect of YQCTF on cell status was detected by Annexin V-PI double staining method.The cell cycle was detected by flow cytometry.The expression of JNK and p-JNK gene was detected by qRT-PCR.2 Animal experiments To construct a human lung squamous cell carcinoma SK-MES-1 cell xenograft model.After successful modeling,they were randomly divided into four groups:model group,Chinese medicine group,chemotherapy group and combination group,respectively,14 days after administration.On the first day,the fifth day,the ninth day,and the 14th day,the body weight and the tumor diameter were observed.After the sacrifice treatment,the tumor weight was calculated,and the pathological observation of the organ tissue was performed,and the JNK and p-JNK proteins in the tumor tissue were taken.Western Blot test.Result:Clinical study1 clinical efficacy:The treatment group(Chinese medicine combined with CCRT treatment)had an ORR rate of 81.4%,a DCR rate of 93.1%,a PFS of 12.5±0.85 months,a control group(CCRT)ORR rate of 74.1%,a DCR rate of 93.1%,and a PFS of 9.2±0.82 months(see Table 3,Table 4).The objective response rate of the tumors in the two groups was tested by rank sum test,and the results were not statistically different(P>0.05).Log-rank test was used to compare PFS differences between two groups of patients,and the results were statistically significant(P?0.05).2 The impact of relevant prognostic factors on PFS:2.1 The effect of TCM syndrome on PFS:The PFS of the treatment group were lung stagnation,spleen deficiency and dampness,yin deficiency and phlegm heat,and qi and yin deficiency were 12.3±1.25 months,14.6±0.16 months,8.6±1.32 months,and 4.2±1.45 months,respectively.The median PFS of the four syndromes were 10.1±0.94 months,12.4±0.70 months,7.9±0.50 months,and 5.8±1.20 months.Log-rank test was used to compare PFS differences among 4 groups of patients,The P values were less than 0.01.2.2 The effect of tumor stage on PFS:The median PFS of patients in stage ?a and ?b of treatment group were 13.7±0.62 months and 8.7±1.94 months,respectively.The median PFS of group ?a and ?b in control group were 10.6±0.57 months and 6.9±0.80 months,respectively.PFS between patients in different stages.Differences were compared by Log-rank test,P values were less than 0.001.2.3 The effect of pre-treatment PS score on PFS:All patients were divided into 3 groups according to PS score 0,1,2 before treatment.The median PFS was 14.60±1.90 months,11.20±0.80 months,8.50±0.59 months,respectively.The difference of PFS between patients in different dose groups was compared by Log-rank test,P=0.005.2.4 Differences in PFS between patients received differentradiotherapy doses:All patients were divided into standard dose group O60Gy)in 85 cases and low-dose group(<60Gy)in 31 cases.The median of standard dose group and low dose group.PFS was 12.40±0.56 months and 7.60±0.56 months,respectively.The difference of PFS between patients in different dose groups was compared by Log-rank test,P value was less than 0.01.3 Cox regression analysis for PFS related factorsNon-genetic factors:gender,T and N staging,objective response of tumor,TCM syndrome,KPS score,myelosuppression,digestive tract adverse reactions were unfavorable factors for PFS,and stage was favorable factors.Genetic factors:PTEN is a favorable factor for PFS,and HER2,PI3K,AKT,mTOR,FGFR,and VEGF were unfavorable factors.4 Differences in and chemoradiotherapy adverse reactions between the two groupsThe independent sample rank sum test(Mann-Whitney U test)was performed on the two groups of patients with bone marrow suppression and gastrointestinal adverse reaction.The results were found to be P=0.090 and P=0.437,both P>0.05.The rate of radiation pneumonitis in both groups was tested by pearson chi-square test,P=0.049<0.05.5 Differences in physiological status,social/family status,emotional status,functional status and additional module scores between the two groups of patients before and after treatment in the quality of life(FACT-L4.0)The scores of the FACT-L4.0 scales in the treatment group were improved compared with those before treatment(P?0.01).According to the classification of TCM syndrome types,except for patients with deficiency of Qi and Yin,other types of patients were treated with FACT.The integrals of each module of the-L4.0 scale were improved compared with those before treatment(P?0.05).In the control group,the scores of the FACT-L4.0 scales were improved compared with those before treatment(P<0.01).According to the TCM syndrome classification,the patients were excluded from the spleen deficiency and dampness type after 5 modules.There was improvement before treatment(P?0.05).The scores of FACT-L4.0 scores in other TCM patients were not significantly improved before treatment(P>0.05).Experimental research:1 Cell experiment The CCK8 method was used to detect the value-added effect of Yiqi Quyu Decoction on SK-MES-1 cells after 48 hours.The Oneway-ANOVA single factor analysis indicated that Yiqi Quyu Recipe could inhibit the increase of SK-MES-1 cells.Concentration-dependent;flow cytometry results showed that different concentrations of YQCTF could induce apoptosis of SK-MES-1 cells in hypoxia with a concentration-dependent effect after one-way analysis of variance with YQCTF for one-hour analysis.Oneway-ANOVA one-way analysis of variance showed that different concentrations of YQCTF could affect the cell cycle,and significantly increased the proportion of cells in G0/G1 phase,and decreased the proportion of cells in S phase and G2/M phase,under the condition of low concentration of YQCTF.The cell cycle can be arrested in the G0/G1 phase,and the cells arrested in the G0/G1 phase increase with the increase of the YQCTF concentration;the q-PCR amplification technique shows that the YQCTF group JNK has no effect after different concentrations of YQCTF.Significant changes,p-JNK expression levels decreased significantly,and had a dose-dependent effect.2 Animal experiments(1)Body weight of nude mice:The weight of the mice in the combined group increased steadily;the body weight of the mice in the Chinese medicine group gradually increased in the pre-and post-dose period,and decreased in the later stage;the weight of the mice in the chemotherapy group was The early stage was gradually decreased,and gradually increased in the middle and late stages;the body weight of the model group mice decreased in the early stage,gradually increased in the middle stage,and continued to decline in the later stage.(2)The effect of tumor growth inhibition rate:The tumor weight of the mice in the Chinese medicine group,the chemotherapy group and the combined group was smaller than that in the model group,and the difference between the groups and the model group was statistically significant(P?0.05).).The tumor weight of the combined group was smaller than that of the traditional Chinese medicine group,and the difference was statistically significant(P?0.05).(3)In terms of tumor inhibition rate:the Chinese medicine group was higher than the model group,but the combined group was higher than the Chinese medicine group and the chemotherapy group.(4)Tumor pathological effects:The tumor cells in the model group were rich in blood vessels,and the cell proliferation antigen was abundant.The other three groups of neovascularization were significantly reduced,the apoptosis of tumor cells was significantly increased,and the proliferation of tumor cells was significantly inhibited.(5)Expression levels of JNK and p-JNK in transplanted tumors:Western blotting results showed that there was no significant change in JNK and significant decrease in p-JNK expression in the YQCTF group compared with the model group,and the dose-dependent effect was significant.(P<0.05).conclusion:Clinical study:YQCTF combined with concurrent chemoradiotherapy for stage III lung squamous cell carcinoma can prolong PFS,reduce the incidence of radiation pneumonitis,improve the quality of life of patients,and improve the efficacy and quality of life among patients with different TCM syndrome types.Experimental research:YQCTF can inhibit the proliferation of human lung squamous cell carcinoma SK-MES-1 cells,arrest the cell cycle,induce SK-MES-1 cell apoptosis,and inhibit the growth of SK-MES-1 cells.Growth inhibition of SK-MES-1 cells may be associated with inhibition of activation of the JNK/SAPK.