The Clinical And Mechanism Study On The Effect Of Yiqiyangyin Decotion(YQYYD) On Prevention For Patients With Early Stage Lung Cancer

Aim of the study: To evaluate the clinical effects of Yiqiyangyin Decotion(YQYYD)on prevention of lung cancer and the inhibitory effect of YQYYD on human lung cancer cell proliferation.To evaluate chemopreventive effect of YQYYD on lung tumorigenesis in A/J mice.Materials and methods: Clinical research: A total of 86 patients with PN(include GGN)were randomized into two groups,treatment group and concurrent control group.The patients in treatment group were treated with YQYYD for 6 months and in both of groups are observed in 2 year.To evaluated the clinical effects by observe the change of PN size and density,quality of life(FACT-L),TCM symptoms and syndromes,PS score and so on.To evaluated the influence on tumor marker and immune function of YQYYD by observe the change of level of CEA,NSE,CD3,CD4,TGF-?,etc.Animal experiments: 40 Female A/J mice were given a twice i.p.dose of B(a)P(100 mg/kg body weight)in 0.2 ml of tricaprylin.Two weeks after 1st B(a)P injection,the mice were randomly divided into 2 groups with 15 mice per group:(a)control group :mice were gavaged distilled water 0.5ml/d;(b)test groups: mice were administered Jinfukang oral liquid(JFK)0.5ml/d by gavage for the duration of the studies.The health condition of the mice was monitored daily and body weights were measured weekly for the duration of treatments.18 week after treatment,mice were sacrificed by CO2 asphyxiation.The fixed lungs were evaluated under a dissecting microscope to obtain fixed surface tumor count and individual tumor size.Tumor volume(V)and tumor burden(N)were measured in each group.Five lung tissues from each group were analyzed.The difference of proliferation and apoptosis protein expression in lung tumor tissue of each groups of mice was detected by IHC.Cell Experiments: Human lung cancer A549(p53-wild type)and H1299(p53-null)cells were t reated with YQYYD of various concentrations.Growth inhibition was detected by MTS metabolism using AQueous one solution cell proliferation assay kits.To investigate protein expression levels of members in the AKT/NF-?B signaling pathways in both of cell lines which were treated with YQYYD,p53 protein expression in A549 cell line by Western blot analyses.The expression of p53 and target genes in A549 cell line was examined through RT-PCR analyses.Results: Clinical research:(1)The change of PN in 6 months and 24 months: The overall curative effect in the treatment group was higher than in the control group,the difference had significant efficiency(P<0.05).(2)Quality of life(FACT-L): After treatment in the treatment group,the physiological status,social/family status,emotional status,functional status,lung cancer specific symptoms in various fields and total scores compared with the control group,there were significant improvement,the difference was significant(P<0.01);In terms of total score,that in the treatment group was higher than in the control group(P<0.01).(3)TCM symptoms and syndromes: In the improvement of the symptoms such as cough,shortness of breath and TCM syndromes,the treatment group was higher than that in the control group(P<0.01,P<0.05).(4)Tumor marker and serum immune function level: The level of NSE was lower after treatment in the treatment group than before treatment,while there was increase in the control group,and the difference wasn't significant(P>0.05).After treatment in the treatment group,the level of TGF-? was significantly lower than before,and the difference was significant(P<0.05).Animal experiments: Treatment with JFK by gavage resulted in a 33% decrease in tumor multiplicity and 46% decrease in tumor burden compared with the control group in A/J mice.Treatment with JFK increased the apoptotic index in mice.Cell Experiment: YQYYD significantly inhibited growth of A549 and H1299 cells in a dose-dependent manner,and were cytotoxic to A549 cells with an estimated IC50 value of 0.22g/ml and to NCI-H1299 cells with an estimated IC50 value of 0.15g/ml after 24 h treatment.YQYYD inhibited phosphorylation of AKT while downregulated the expression of phospho-IKK?/?.The level of phospho-NF-?B and phospho-AKT were dramatically decreased after YQYYD treatment 24 hours in A549 cells.The expression levels of proteins were no significantly changed in H1299 cell line.Exposing A549 cells to YQYYD for 24 h noticeably increased p53 protein expression in a time-dependent manner.RT-q PCR revealed that YQYYD treatment upregulates m RNA levels of proapoptotic p53 and target genes(Bcl-2 family genes)BAX,Bim and Puma in A549 cells.Conclusions:(1)YQYYD has the effect of delaying or preventing PN canceration and preventing the occurrence of lung cancer.(2)YQYYD can improve the quality of life and relieve the symptoms of patients with PN.(3)YQYYD can play the preventive role of lung cancer by reducing the level of TGF-?.(4)YQYYD could inhibit the proliferation of lung cancer cell through p53 dependent AKT/NF-?B pathway in vitro.(5)Yiqiyangyin formula has the chemopreventive effect on lung tumorigenesis in mice.(6)TCM Yiqiyangyin principle can be used as one of the effective treatments on the prevention of lung cancer.