Study On Inhibiting Lung Carcinoma A549 Cell And Its Targets Of Serum Containing Taxus Chinensis Var.Mairei(AETC)

Objective To study the proliferative effect of serum containing Taxus chinensis var.mairei(AETC)on human lung carcinoma cell A549 and its targets such as EGFR,so as to explore the function.Methods Eight purebred New Zealand rabbits,weighing about 2000g,were randomly divided into four groups:A group:the control group treated with distilled water,B group:the low dosage group treated with 186.65mg/kg/d AETC,C group:the moderate dosage group treated with 373.3mg/kg/d,D group:the high dosage group treated with 746.6mg/kg/d AETC.All the treatments were administered for 7 successive days intracardially.Rabbits’ blood serum was harvested via cardiac puncture 1 h after the last administration.All the serum in one group were mixed and placed at 56℃ for 30 min for inactivating complement,then filtered through 0.22 μm micropore filtrate for sterilization and conserved at-20 ℃ until use.The drug-containing serum is diluted with F12k solution as 10%AETC pharmacological serum.After Human lung carcinoma cell A549 is cultivated 24 hours,we detect OD value with MTT assay,and calculate inhibition ratio.The express of protein of EGFR.pEGFR is examined by Elisa and Western blot,and the gene expression of EGFR、VEGF mRNA is examined by RT-PCR.All data are expressed as mean ± standard deviation(SD)of three experiments.All the statistical analysis were performed by SPSS 17.0 software.Statistical significance was compared between each treatment groups and control group by one-way ANOVA.Values with P<0.05 were considered as statistically significant.Results MTT assay revealed that compared with the control group,serum containing AETC groups significantly decreased the viability of A549 cell,showing a dose-and time-dependent manner.Elisa method showed that compared with the control group,the moderate dosage group decreases the expression of EGFR and VEGF(P<0.05).Western blot method showed that serum containing various concentrations of AETC treatment strongly reduced the level of phospho-EGFR(P<0.05).Real-time RT-PCR shows that serum containing various concentrations of AETC treatment strongly reduced the level of EGFR mRNA(P<0.05)and VEGF mRNA(P<0.05).Conclusion Our study demonstrates that serum containing AETC inhibits the proliferation of A549 cells by suppressing the targets such as EGFR and VEGF.