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To observe the effect of Yiqi Chutan Tang on Lewis lung cancer tumor growth in C57 BL mice and protein screening.Lung cancer tumor model was established in C57 BL mice.40 mice were randomly divided into model group and treatment group.Tumor size,tumor weight and the numbers of lung metastases were detected.The expressed differentially protein spots information were acquired by a two-way fluorescence difference gel electrophoresis (2D-DIGE) system,and the differential proteins were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-TOF).The differences in protein expression was finally verified by Western blot and fluorescence quantitative PCR.We found that Yiqi Tumor size,tumor weight in Chutan Tang treatment group were significantly less than that in model group (p<0.01),and the tumor growth inhibition rate was 57.21%.For gel diagram analysis of 2D-DIGE system,compared with model group,there were 44 expressed differentially protein spots,of which 6 protein spots were up-regulated and 38 protein spots were down-regulated in the Yiqi Chutan Tang treatment group.Among these proteins,37 proteins including 30 down-regulated proteins and 7 up-regulated proteins were successfully identified by MALDI-TOF-TOF.Hsp-60,P4HB,PDI-A3 and EG433182,Hsp5p,Hsp9,STIP1 were verified down-regulated respectively by Western blotting and qPCR in treatment group.In conclusion,The possible mechanism of Yiqi Chutan Tang in the treatment of LEWIS lung cancer was highly related with the down-regulated protein expression of Hspd1,prolyl 4-hydroxylase,protein disulfide-isomerase A3 precursor,EG433182,heat shock protein 5 precursor,heat shock protein 9,stress-induced phosphoprotein 1.