| ObjectivesThis in vitro study was to observe the influence of curcumin on human esophageal cancer cell line EC9706, and whether it has the function as proliferation or apoptosis on EC9706 and its effects on the invasion and proliferation of esophageal cancer cell line EC9706, and to explore its molecular mechanism which aims to lay a good foundation for the further study of its extensional mechanisms as antitumor.Materials and methods1.The experiment of cell proliferation and apoptosis(the study of curcumin's influence on esophageal cancer cells in vitro), with the method MTT. Divided them into three groups when the cells adherent after adjusts the cell concentration of EC9706 which is during logarithmic growth phase to 1×105/mL (Routinely cultured cells of human esophageal carcinoma cell lines EC9706):①the Experimental group: added curcumin which's final concentration was 40μmol/L,80μmol/L,120μmol/L and 160μmol/L into cells;②the Negative control group:added the culture medium RPMI-1640 into cells;③the Solvent control group:DMSO was administered to culture tumor cells with 0.1%. Set 12 duplicate wells in each group, respectively curcumin intervention 24h,48h,72h, to detect the differences of each group in cell proliferation inhibition rate.2. To detect the change of expression of Ets-1 and MMP 9 mRNA before and after the curcumin's treatment by RT-PCR. Routinely cultured cells of human esophageal carcinoma cell EC9706, adds the culture medium RPMI-1640 into the control group, and added curcumin of the final concentration which wss 40μmol/L, 80μmol/L,120μmol/L and 160μmol/L into the experimental group.Respectively acting on the cell after 48h and 72h. Collecting 3 bottles of EC9706 cells of the control group and the experimental group to detect the expression of ETS-1 and MMP9 mRNA by RT-PCR.3. To detect the influence of curcumin on the protein expression of EC9706 ETS-1 and MMP9 with immunohistochemical methods.4. To analyze the data by SPSS 13.0 statistically, to test the statistical analysis of rates byχ2, to analyze the comparison among groups by single factor variance. It has the statistical significance when P<0.05 as the test standard of a=0.05.Results1. MTT:The results showed that the curcumin had the function as proliferation and Inhibition on EC9706, and it was line-depended on the concentration and time. In addition the differences were great (P<0.05). After 24h,48h and 72 hours with the treatment of curcumin, the IC50 were 206.3μmol/L,135.3μmol/L and 98.4μmol/ L,respectively. After 72 hours'treatment of 160μmol/L curcumin,EC9706 cells' proliferation almost stopped.2. RT-PCR showed the expression of Ets-1 and MMP9 mRNA in the EC9706 cells which were treated by the curcumin decreased while the concentration gradually increased. At the same time, the expression of them gradually decreased following time with the effect of the 120μmol/L curcumin.3. After the treatment of the curcumin, the protein expression of Ets-1 and MMP9 in EC9706 cells both decreased, and there was relevance with statistical significance. (r=0.914,P<0.05)Conclusions1. Curcumin can inhibit the proliferation of human esophageal cancer EC9706 cells effectively while by inducing the apoptosis of the cells, and the function is positively correlated with dose and time. This process may be related to the expression of the curcumin inhibition of Ets-1 and MMP9 gene.2. Curcumin can inhibit the protein expression of Ets-1 and MMP9 in EC9706 cells, relativity may exist between the expression of Ets-1 and MMP9. This may be one of the mechanisms by which curcumin inhibit the apoptosis of tumor cell. |
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