To Study Gene Expression Pofiles Differences Between The Rat Bone Marrow Mesenchymal Stem Cells And Differentiated Liver Cells

Gene chip has the advantages of high efficiency, high flux, high precision, so it was quickly applied to animals, plants and human genes。To analysis gene expression profile by sequenced high throughputly on a species or specific cells in the mRNA from a functional state. Through analysising of the genes express change, it revealed the relationships between the changes of gene expression and achieved biological information.[objective]In this study, we analysis the differences of gene expression among the three groups of the rat bone marrow mesenchymal stem cells, hepatocyte like cells, normal liver cells. wanted to research the express trend clustering and function analysis because of the differential gene during the process of bone marrow mesenchymal stem cells into liver cells.To used the PCR technology to verify significant difference gene, and wanted to find the nuclear mechanism possibly to supply information for clinical application.[method] 1.After compared with the differentiation of rat bone marrow mesenchymal stem cells and hepatocyte like cells, we screend the differences of gene expression among the two. To analysis of function about the difference genes and significance analysis about pathway among bone marrow mesenchymal stem cells and hepatocyte like cells, we aquired significant function gene and Pathway among differential expression genes with GO and pathway analysis. Through constructed of differential gene expression regulation network,we acquired the core genes and pathways about the differentiation of liver cells.2.After compared with the differentiation of Hepatocyte like cells from rat bone marrow mesenchymal stem cells and Normal liver cells, we screend the differences of gene expression among the two. To analysis of the the difference about significance function and Pathway among hepatocyte like cells and Normal liver cells, we aquired significant function gene and Pathway among differential expression genes, we aquired the significant Pathway and function involved differences with GO and pathway analysis. Constructing gene regulatory network through those differential gene expression and according to expression abundance (express the signal value) in the network and compary the networks between HLCs and NLCs, acquired the core of genes and pathways of the differentiation of liver cells.3. In rat bone marrow mesenchymal stem cells, induced differentiation of hepatocyte-like cells, normal liver cells for the study of the development process, we screened differentially expressed as a dynamic process.are differentially expressed genes among three sets. Using differential gene expression of three cell states and according to the expression of the dynamic change process clustering trendsUsing the differentially expressed genes of three group cells, we screened significant expression of the trend as the control of bone marrow mesenchymal stem cells into hepatocyte gene. To Analysis with the same trend of co-expressed genes and predict whether a gene co-expression is regulated by transcription factors which appeared showing consistency in expression. We constructioned expression dynamic process of gene regulatory networks, screened core genes in the dynamic process and according to the interaction between genes, we get in the regulatory mechanism of the dynamic process.4. In order to confirm the results of the chip reliability, We used fluorescence quantitative PCR experiments related genes with further chip verification experiment.[Results]1. By screening we can get 839 differentially expressed genes between bone marrow mesenchymal stem cells and differentiated hepatocyte-like cells, among 448 genes up-regulated,391 genes down-regulated.The Pathway analysis results revealed 45 pathways had significant changed among the 273 Pathways,275 differentially expressed genes involved in those pathway, including 122 up-regulated genes and 153 down regulated genes. In the network, we selected 5 key genes:Pkm2, Nt5e, Fos, Adcy3, Itga7.2. We can get 4171 differentially expressed genes between normal liver cells and induce differentiation of hepatocyte-like cells,among 1804 genes up-regulated,2367 genes down-regulated.Pathway analysis results revealed 699 pathways had significant changed among the 699 Pathways,including 344 up-regulated genes and 355 down regulated Genes.3. Among the dynamic process from MSCs differentiated into NLCs, we can get 4938 differentially expressed genes and Selected 7 key genes:Fas> G6pc. Il1b、S100a1、Ndufa7, etc.4. Real-time PCR results were validated analysis combined with global gene expression analysis of GO analysis and Pathway credible selected high value, In MSCs HLCs genes induced further chip verification experiment, we verified the reliability of the results of the chip.[conclusion]This procedure that MSCs was eventually differentiated into HLCs are closely related these regulate genes expression just as Cell cycle regulation, proliferation and differentiation and associated with liver cell function as:Sugar, fatty acid, cholesterol metabolism, detoxification.