| ObjectiveInvasion of trophoblastic cell to endometrium is essential for implantation and early embryo devepment. This process depend on precise regulation of multiple genes expression. In this study, we have analysized gene expression profiles by gene array technology, and probe molecular basis of EVT invasion in abnormal pregnancy. It is important for illumination of pathogenisis and mechanisim of gene network regulation.Methods1. CTB and EVT of villous and decidua from spontaneous abortion during pregnancy 8 w to 12 w were Collected by trypsin digestion method.2. Total RNA was extracted from CTB and EVT respectively, Then detected RNA purity and integrity.3. Gene chip hybridization and gene expression profiles data extraction were performed in Shanghai Biochip Limited Company.4. Differential expression genes were obtained by analysis of gene expression profiles data .5. Selected two differential expression genes, KISS1 and MMP-10, and detected their expression by RT-PCR and immunohistochemistry to confirm confidence of gene chip data.6. Analysis linking to metabolic pathway, different biology functions of differential expression genes were analysized.Results1. By analysis of RNA purity and integrity, A260/280 is 1.9 to 2.1. Eletrophoresis bands of 28sRNA and 18sRNA are very clear. Extracted RNA accord with the experimental requirement2. High purity CTB and EVT were collected by the two different concentration trypsin digestion methods.3. The gene expression profiles chip was founded, which quality control was up to standard. The result of analysizing to KISS1 and MMP-10 gene expression by RT-PCR and immunohistochemistry were coincidence with that of analysizing by gene chip, which indicated gene chip data were confidence.4. Among gene expression profiles of CTB and EVT, 2308 differential expression genes were found. Cluster analysis showed that there were 933 genes up-regulated and 1375 genes down-regulated in theses differential expression genes. The biological function of all the genes were queried and classified according to the standard of GeneOntoloty. We analysed 2308 differential expression genes, the result showed that 307 genes were up-regulated and 391 genes were down-regulated in genes related to metabolism, 29 genes were up-regulated and 5 genes were down-regulated in genes related to extracellular matrix, 54 genes were up-regulated and 153 genes were down-regulated in genes related to signal transduction, 87 genes were up-regulated and 58 genes were down-regulated in genes related to transcription, 36 genes were up-regulated and 78 genes were down-regulated in genes related to cell growth, 13 genes were up-regulated and 41 genes were down-regulated in genes related to apoptosis, 62 genes were up-regulated and 80 genes were down-regulated in genes related to transportion, 30 genes were up-regulated and 53 genes were down-regulated in genes related to stress reaction, 12 genes were up-regulated and 112 genes were down-regulated in genes related to immune response, 37 genes were up-regulated and 53 genes were down-regulated in genes related to cell adhesion, 58 genes were up-regulated and 55 genes were down-regulated in genes related to development. In addition, the function of 539 genes had not been described on GeneOntoloty at present , 209 genes of them were up-regulated and 296 genes were down-regulated. ConclusionsCTB and EVT were analysized by the globe gene expression profiles chip. 539 genes of the 2308 differential expression genes (or EST) had not been biological function described on GeneOntoloty at present. Meanwhile, some genes related spontaneous abortion were confirmed in this study.By analysis to differential expression genes, We suggestied there be following regulatory mechanism in process of trophoblasts invasion. 1. Comparating to normal pregnancy, genes related to cell adhesion and extracellular matrix were down-regulated, it indicate that change of these genes expression may influence recognition, adhesion, degradation of extracellular matrix, re-establishment of uterus blood vessele and result in abortion. 2. In deciduas with spontaneous abortion, lower expression of the genes related to immune response and defense response function may induce immunological rejection of mother to embryo and it may be an important cause for spontaneous abortion. 3. Expression change of genes related to signal transduction and cell cycle regulation function may promote cells to proliferated and differentiated in early embryo development. 4. The intensity expression of genes related to cell growth and development function were worthiness to maintain embryo cells growth and formation of organs. 5. Compared to gene expression profile chip of normal pregnancy, The number of up-regulated genes related to transportation and metabolism function were less than the number of down-regulated genes in spontaneous abotion. The cause may be relate with change of pathology. |
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