Serum Containing AETC Inhibit Lung Carcinoma A549 Cell Proliferation Through MAPK Pathway

Objective:To study the proliferative effect of serum pharmacological Aqueous Extract of Taxus chinensis var.mairei(AETC)on human lung carcinoma cell A549,and to explore the effect of signal pathway Mitogen-activated Protein Kinase(MAPK).Method: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?m 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 took pictures under microscope and detect OD value with MTT assay,and calculate inhibition ratio.The express of protein of ERK?P-ERK?p38?P-p38?JNK?P-JNK is examined by Western-blot,and the gene expression of ERK1?JNK?p38 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.Result:MTT assay revealed that compared with the control group,serum containing AETC groups significantly decreased the viability of A549 cell,the inhibition ratio of the three trial groups is:B group:19.73%,C group:32.46%,D group:42.23%.Western blot method showed that serum containing various concentrations of AETC treatment strongly reduced the level of phospho-JNK and phospho-ERK1/2 while increased phospho-p38.However,no significant effects on the expression of JNK,ERK1/2 and p38 MAPK were detected in our works.Real-time RT-PCR shows that serum containing various concentrations of AETC treatment strongly reduced the level of ERK1?JNK mRNA(P<0.05)while increased p3 8 mRNA(P<0.05).Conclusion:Our study demonstrates that serum containing AETC inhibits the proliferation of A549 cells by suppressing the MAPK signaling pathway.