Study Of The Effect Of Aqueous Extract Of Taxus Chinensis On Human Lung A549 Cell And Which Combined With Erlotinib EGFR Expression

Study of the Effect of Aqueous extract of Taxus chinensis on human lung A549 Cell and which combined with Erlotinib EGFR expressionObjective To study the effects of Aqueous extract of Taxus chinensis on PCNA, VEGF and MAPK pathway expression and the possible mechanism from PCNA, angiogenesis and signaling pathway on protein level. To study the inhibitory effect of aqueous extract of T.chinensis var combined with Erlotinib on vitro cultured human lung adenocarcinoma A549 cell, and effects of EGFR expression. It provids experimental basis for clinical treatment of Aqueous extract of T.chinensis var to Lung cancer.Methods①The level of PCNA, VEGF and MAPK pathway in vitro cultured human lung adenocarcinoma A549 cell were detected by ELISA and Western blot after the affects for 24 hours. Detecte the expression of protein PCNA,VEGF,ERK,P-ERK,P38 and P-P38 after Aqueous extract of Taxuschinensis of three different densities 2.0mg/ml,2.5mg/ml and 3.0mg/ml and blank control group functioning on cell line A549 24h later.②Screening the optimal concentration of Erlotinibt for Inhibit human lung adenocarcinoma A549 by MTT method. MTT assay was used to observe the anti-proliferation effect of T.chinensis var extract combined with Erlotinib on A549 cells for 24 h. Western blot assay was used to detect the EGFR,P-EGFR expression on A549 cells.20μM Erlotinib combined with three different densities of Aqueous extract of T.chinensis var 2.0mg/ml,2.5mg/ml,3.0mg/ml culture A549 cell for 24h.③MTT and Webstern-blot assay was used to observe the anti-proliferation effect of T.chinensis var extract combined with Erlotinib on A549 cells for 24 h. A549 cell were divided into four groups,20μM Erlotinib combined with three different densities of Aqueous extract of T.chinensis var 2.0mg/ml(Ia),2.5mg/ml(Ib),3.0mg/ml(Ic); 20μM Erlotinib; 2.5mg/ml Aqueous extract of T.chinensis var and blank cont rol(Ⅳ).Result①The best inhibition rate of Erlotinib is when rate 20mM,and its inhibition rate is 27.87%.②The inhibitory rates at the combined groups were 54.330%,61.96%,71.84% respectively, showed the statistical significance compared with the Eltinib group(27.8%)and the DMSO group(P<0.05 or P<0.01).③Aqueous extract of Taxuschinensis can down-regulate expression of protein PCNA,P-EGFR, P-ERK.of A549 cell.④Aqueous extract of Taxuschinensis can down-regulate expression of protein P-EGFR in cell line A549.After combined with Erlotinb,expression of protein P-EGFR obviously decreased.Conclusions①Aqueous extract of Taxuschinensis can influence MAPK signal pathway and up-regulating P-P38 down-regulating P-ERK,PCNA maybe one of mechanisms of its antitumor function.②The effect of increasing A549 cell inhibition on Aqueous extract of Taxuschinensis combined with Erlotinib but whether have the synergistic effect needs to further research.③Aqueous extract of Taxuschinensis can down-regulating P-EGFR protein maybe one of mechanisms of its antitumor function.