The Study Of Relation Between The Drug Resistance Of EGFR-TKI And The Cold Or Heat Syndrome Type In Patients With Stage â…¢B/â…£ NSCLC

Objective1.Clinical study:By observing the characteristics of cold and heat syndrome type of stage â…¢B/IV NSCLC patients who treated with EGFR-TKI, especially in the drug resistance moment, to verify the property of EGFR-TKI, discuss the relation between the cold and heat syndrome type and the therapeutic effect of EGFR-TKI. For further study, providing theoretical foundation for researching the reversion of drug-resistance by TCM.2. In vitro study:By observing the effect on cell proliferation and apoptosis test of different therapeutic principle of TCM(clearing heat or warming yang) combined with Gefitinib, and exploring the mechanism, to screen which therapeutic principle of TCM can increase sensitivity towards Gefitinib and verify the property of EGFR-TKI. For further study, providing theoretical foundation for TCM increasing sensitivity towards EGFR-TKI and clinical trial.Methods1. Clinical study:Applying observational epidemiological method, this study enrolled 138 stage â…¢B/â…£ NSCLC patients who were preliminary diagnosed without prior anticancer treatment, or those recurred with an interval more than 6 months from last treatment. All were scheduled to be treated with EGFR-TKI. Collected their four TCM diagnostic methods and identified cold or heat syndrome type by TCM theory. The endpoint was drug-resistant moment(PFS). Established a database to discuss the changes of cold and heat syndrome type characteristics during the treatment of EGFR-TKI, especially the TCM syndrome type in the drug resistance moment.2.In vitro study:Human lung adenocarcinoma cell lines A549,H1650 were the object of this study. Warming yang Chinese patent medicine combined with Gefitinib and clearing heat Chinese patent medicine combined with Gefitinib were added to the cells lines respectively. MTT was used to test the inhibition of the cell proliferation which were cultured by the above drugs. Flow cytometry was used to test the apoptosis of the cells after cultured by the above drugs.Western blot was used to detect the mechanism of action.Result1. Clinical study:(1) The differences in smoking history and pathological type of patients were statistically significant (P<0.05).(2) The overall analysis showed that the TCM syndrome types of most patients would change from yin-cold to yang-heat after the treatment of EGFR-TKI gradually. Before treatment,31.4% patients were with yang-heat syndrome types,68.6% patients were with yin-cold syndrome types. As time went on, the proportion of patients with yin-cold syndrome declined and the proportion of patients of yang-heat syndrome risen. In the drug-resistance moment, majority of the patients were with yang-heat syndrome types(76.2%).(3) Stratified analysis showed that the difference between resistance syndromes with the syndromes before treatment and the syndromes after 1 months treatment were no statistically significant (P<0.05). The difference between resistance syndromes with the syndromes after 3 or 6 months treatment were no statistically significant (P>0.05)(4) The age, smoking history and gender has no statistically significant with the resistant time (P>0.05).(5)The median PFS of yang-heat syndrome patients were 1.9 months, yin-cold syndrome patients were 5.267 months. The TCM syndrome types before treatment has no statistically significant with the curative effect(PFS) (P>0.05)2. In vitro study:(1) A549:There was no interaction effect between Gefitinib and ShenFu injection from MTT test(P>0.05). The Q value showed the two drugs had additive effect (0.85<Q<1.15).The apoptosis rate had no statistically significant difference (P>0.05).There was interaction effect between Gefitinib and QingKaiLin injection from MTT test (P<0.05). The Q value showed the two drugs only had additive effect (0.85<Q<1.15).The apoptosis rate had risen after the treatment with the combination of the two drugs (P<0.05), but the mechanism of action was not found.(2) H1650:There was no interaction effect between Gefitinib and ShenFu injection from MTT test (P>0.05).The Q value showed the two drugs changed from additive effect to antagonism 48H:0.85<Q<1.15,72H:Q<0.85). The apoptosis rate had no statistically significant difference (P>0.05).There was interaction effect between Gefitinib and QingKaiLin injection from MTT test (P<0.05). The Q value showed the two drugs had synergistic effect (Q>1.15). The apoptosis rate had risen after the treatment with the combination of the two drugs (P<0.05).The mechanism of action may via downgrading the expression of EGFR and p-ERK (P<0.05)Conclusion1.As time went on, the proportion of patients with yin-cold syndrome declined and the proportion of patients of yang-heat syndrome risen after the treatment of EGFR-TKI. In the drug-resistance moment, most patients were with yang-heat syndrome types. So that the property of EGFR-TKI may be yang-heat nature.2. There was no statistically significant difference between the TCM syndromes types pre-treatment and the curative effect(PFS). But the yin-cold syndrome patients had benefit tendency.3.To A549, there was additive effect between Gefitinib and ShenFu injection. The combination of Gefitinib and QingKaiLin could rise the inhibition ratio and apoptosis rate. The signaling pathway of action was not found.4. To H1650, there was additive effect between Gefitinib and ShenFu injection. The combination of Gefitinib and QingKaiLin could rise the inhibition ratio and apoptosis rate obviously. The signaling pathway of action may via downgrading the expression of EGFR and p-ERK.