The Effect Of Pokemon On Gene Expression Profile In MCF-7 Breast Cancer Cell

[Objective]The gene microarray and bioinformatics technology were used to research the effect of Pokemon on gene expression in MCF-7 breast cancer cell, and to discuss the effect of Pokemon on cell biological function.[Methods]The experiments included two parts. In the first part, there were two kinds of MCF-7 breast cancer cell. The treatment group was stable over-expression of Pokemon and the control group was stable expression of Pokemon. The total RNA of two groups was extacted respectively and then the gene microarray was used to detect the gene expression profile of the two samples. Next, the acquired microarray datas were analyzed through clustering analysis, Pathway analysis and GO analysis, etc. And Real-time PCR technology was used to vertify the result of the gene expression profile. To vertify the result above, the Real-time PCR technology was used again to detect the expression of TGF-β1 and SMAD4 in the MCF-7 cell with transient expression of Pokemon. In the second part, tissue chip and immunohistochemistry staining were performed to evaluate the expression of TGF-β1 in 208 cases consisting of breast carcinoma tissues and the edged tissues in corresponding, and to analyze the expression of TGF-β1 expression in different tissues.[Result]1.The amount of total genes which expressed differentially in MCF-7 breast cancer cell with stable over-expression of Pokemon was 2079 compared to the control group(the fold change >2.0). There were 966 up-regulated genes including AKT1, WASF2, etc, and 1113 down-regulated genes including TGF-β1,SMAD4, etc. All in all, the number of down-regulated genes was more than the number of up-regulated genes. We knew that the ability of cell proliferation had been decreased after transfected the Pokemon through clustering analysis. 2.Through the pathway analysis, we found that the number of down-regulated genes was more than the number of up-regulated genes in participating in the immune system, adaptive immune system, fatty acid, triacylglycerol, ketone body metabolism, cancer, P53 signaling pathway. However, the number of up-regulated genes was more than the number of down-regulated genes in participating in the metabolism of lipid and lipoproteins, hemostasis, developmental biology, MAPK signaling pathway. 3.Through the GO analysis, mainly from the next three aspects that were cellular component, molecular function and biological process, we found that the down-regulated genes dominated in the most of the aspects.4.The result of the gene expression profile was been vertified by the Real-time PCR technology.5.The result of the gene expression profile was been vertified again through the Real-time PCR technology, that detected the expression of TGF-β1 and SMAD4 in the MCF-7 cell of transient transfected Pokemon plasmid. 6.The expression of TGF-β1 was higher in breast carcinoma than in the edged tissues in corresponding, and there was statistically significant difference in this comparison(p<0.05).7.In breast carcinoma, the expression of TGF-β1 had no significant correlation with ages, tumor types, nuclear grades and node metastasis(p>0.05).[Conclusion]1.The regulation of Pokemon on the expression of genes correlated with cell proliferation and metastasis is mainly nagative, and the regulation of Pokemon on the expession of genes correlated with cell differentiation is mainly positive in the MCF-7 breast cancer cell. 2.Pokemon can downregulate the expression of TGF-β1 and SMAD4. 3.TGF-β1 is over-expression in breast carcinoma, and the expression of TGF-β1 had no significant correlation with ages, tumor types, nuclear grades and node metastasis.