The Study Of Evodiamine To Interfere The Glycometabolism Of Tumous

ObjectivesEvodiamine (Evo) is the effective constituent of the traditional Chinese medicine,which is fructus evodiae. Studies have proved that Evodiamine has stronger anti-tumor effect and the effective concentration is low. It can induce the apoptosis effect for a wide variety of tumor cells. By gene chip research findings, Evodiamine can Promote the apoptosis of tumor cells by inhibiting the expression of multiple pathways.The effect related to the pathways of specific glucose metabolism and Warburg effect in tumor.In the meanwhile, Evodiamine have an effect on some key enzymes in glucose metabolism pathways.The research will give priority to medicine of Evodiamine to explore the mechanism of the pathways of specific glucose metabolism in tumor.Method1、Using Real-Time qPCR technology to explore different Time points of tumour-specific glycometabolism pathways and key enzymes,and apoptosis pathway. the effects included the gene expression of HIF1-a, Akt, PFK, PRS1 and caspase 9;2、Using the technique of Western blot to detect on the expression by key enzyme protein of glycometabolism and the apoptosis protein, including the PFK,RR,p53 and caspase9;3、Using the determined by MTT method to detect the combination effect of evodiamine with paclitaxel on human liver cell HepG2 synergies;4、Using the Flow cytometry to detect the evodiamine and paclitaxel on apoptosis of HepG2,detect evodiamine and paclitaxel combination of apoptosis;5、Tumor-burdened mice to investigate the evodiamine and taxol inhibition in mice liver metastases, set up the normal control group, model group, the 5-Fu positive group of 10 mg/kg, evodiamine of 40 mg/kg group, evodiamine of 80 mg/kg group to observe tumor growth and inhibition.Results一、Technical inspection of the Real-Time qPCR evodiamine to HIF1-α, PI3K-PKB/Akt, PFK, PRS1 role.By evodiamine effect on liver cancer cell HepG20h,12 h,24 h after testing HIF1-a and PI3K-PKB/Akt, PFK, PRS1 gene expression, it is found that evodiamine effect after 12 h, HIF1-α, PI3K/PKB/Akt, PFK, relative sample template PRS1 gene were 0.42,0.60,0.65,0.54; Evodiamine effect after 24 h, HIF1-α and PI3K/PKB/Akt, PFK, PRS1 gene relative sample templates are 0.49,0.44,0.51,0.53, respectively. Above shows that alkali drug under the action of evodia rutaecarpa can cut more than the expression of genes, evodiamine have inhibition of sugar metabolism related pathways and the expression of metabolic enzymes.二、Western blot detecting evodia base on sugar metabolism related enzyme protein expressionIn evodiamine effect on liver cancer cell HepG20 h, by contrast, testing PFK,RRM2,aldolase,p53 and caspase9 protein expression, it is found that 12 h PFK protein expression compared with control group, all have inhibition, explain evodiamine expression of related glycolytic enzyme inhibition, thus inhibiting tumor growth.三、evodiamine base joint of paclitaxel on human liver cell HepG2 synergies effect evodiamine in different concentration of alkali and different concentrations of paclitaxel combination effect of observation on liver cancer cell HepG2,0.3 umol/L evodiamine joint 0.005 umol/L paclitaxel,0.3 umol/L evodiamine joint 0.01 umol/L paclitaxel,0.6 umol/L evodiamine joint 0.005 umol/L paclitaxel,0.6 umol/L evodiamine joint 0.01 umol/L paclitaxel,1.2umol/L evodiamine joint 0.005 umol/L paclitaxel,1.2 umol/L evodiamine joint 0.01 umol/L paclitaxel on HepG2 cell inhibition rate were 30%±0.002,40%±0.007,42% ±0.02,49%±0.01,46%±0.04, 51.3%±0.03 jung-kun Kim Q values were 1.58,1.13,1.85,1.58, by the result, the low concentration of evodiamine and low concentrations of paclitaxel combination of hepatoma cell HepG2 have the function of the synergies, almost all the Q value is greater than 1.15, in addition to 1.2 umol/L evodiamine joint 0.01 umol/L drug taxol group Q value was 1.00.0, Flow cytometry detection of evodiamine and paclitaxel on HepG2 cell apoptosis.By flow cytometry detection, found evodiamine with single drug taxol on apoptosis cells can produce effect, and the drug combination of apoptosis effect enhance, evodiamine and paclitaxel combination effect and promote apoptosis of HepG2 fine effect.五、To control the growth of tumour which in the interal of the mice using the EvoAfter inoculated with H22 liver cancer cells in mice, by setting the model group, the group of 5-Fu by 10 mg/kg, evodiamine by 40 mg/kg, evodiamine by 40 mg/kg to observe the tumor growth inhibition. Results found that 5-Fu by 10 mg/kg and evodiamine by 40 mg/kg have different levels of tumor inhibition compared with the model group; And evodiamine 80 mg/kg group tumor has no inhibition, the reason may be that bad instead absorb high concentrations of drugs, in the body can’t play a role of inhibition, it is possible that high concentrations of insoluble drug, also can’t lead to display the good effect.ConclusionEvodiamine has stronger antitumor, it can inhibit some specific pathway of glycometabolism in tumor.research found that evodiamine can inhibit some key enzymes in the glycometabolism pathways and Warburg effect. Combined the Evodiamine and paclitaxel can induce the cancer cells of HepG2 to die.