Bioinformatics Mining From The Hypoxic Treatment Of Human Arterial Endothelial Cell Sage Library Hypoxic Response Related Gene,

Hypoxia is important component of many physiological and pathological processes. In China, diseases that cause the highest death rate, such as cardiopathy, apoplexy, tumor and respiration disease are all directly or indirectly related to hypoxia. It is very significant to find out the mechanism of how organism responses to hypoxia. In this study, we utilized bioinformatics method to screen the Serial Analysis of Gene Expression(SAGE) library constructed from human aortic endothelial cells(HAECs) exposed to short-term chronic hypoxia and dig out many novel genes related to hypoxia response. We also utilized 3D-PSSM and CDD (Conserved Domain Database) to predicted the function of their protein products and did further bioinformatical analysis to the interested novel genes. Finally, we cloned and checked the expression of two of the novel genes. The results are as follows.1. Find out 998 novel genes related to hypoxia response from 40629 tags (12289 unique) derived from the Serial Analysis of Gene Expression (SAGE) library constructed from human aortic endothelial cells (HAECs) exposed to short-term chronic hypoxia.2. Find out 369 novel genes whose protein products are predicted functional by 3D-PSSM and CDD from 998 novel genes related to hypoxia response. We classified them to 14 clusters. Most of them were related to signal transduction, gene expression regulation, cytoskeleton, or post-translation modification.3. Selected 8 interested novel genes to do further bioinformatical analysis and picked out gene1 and gene4 to clone.4. Constructed the eukaryotic expression vector (pcDNA3.1/Myc-His(-) A MCS) of mouse homolog of gene1; constructed the eukaryotic expression vectors (pcDNA3.1/Myc-His(-) A MCS) of human sequence and mouse homolog of gene4. The genes were proved to be correct by sequencing.5. Using semi-quantitative RT-PCR method, the changes of expression level of gene4 was also confirmed in hypoxia treated human umbilical vein endothelial cells (HUVECs).