人脐静脉内皮细胞基因表达谱分析
摘要
研究背景：
内皮细胞在人类许多重大疾病发生过程中处于重要的地位，例如冠心病、肿瘤等。冠心病是由于冠脉血管病变导致心肌细胞缺血、缺氧，使该支冠脉血管(包括微循环)及所支配区域的心肌细胞凋亡、坏死，其中内皮细胞的的应激、炎症反应、脂质沉积等病理生理过程是冠心病发生的重要机制。在肿瘤尤其是实体瘤的发生过程中，血管新生是肿瘤得以存活的重要因素，而血管新生的最主要过程即为内皮细胞的生长，另外肿瘤的侵袭、转移等过程，也有赖于内皮细胞的参与。因此可以看出，内皮细胞在这些关乎人民健康的重大疾病的发生过程中占有重要的地位。
基因在疾病的发生发展过程中处于重要的地位已成为科学界的共识。基因表达的改变在内皮细胞病变的过程中处于什么地位?发挥什么作用?——长期以来就是医学领域中研究的重点和热点。当前，对于内皮细胞基因表达方面的研究较多，但是尚未见将内皮细胞全部基因表达谱进行全部分析的报道。若能将内皮细胞的所有表达基因进行分析，汇总和分类，分析这些表达基因所参与的病理生理功能及在内皮细胞中所发挥的作用，那将对基因治疗运用于内皮细胞具有重要的意义。
目的：
利用当前基因芯片高通量技术，全面分析人脐静脉内皮细胞的基因表达谱，对所有表达的基因进行生物信息学分析，汇总整理出与内皮细胞相关的重要的基因、分子功能和信号通路。
方法：
(1) 应用人类HG-U133A plus 2.0芯片对人脐静脉内皮细胞的基因表达谱进行全面分析，进而就用荧光定量PCR方法对结果进行验证。
(2) 应用Microsoft Excel、Milano、Go、PANTHER以及Genomatix等软件和网站对基因表达谱进行分析、汇总及数据挖掘。
Introduction
Endothelial cells (ECs) are very important for many diseases, for example, coronary artery disease, and tumor. Coronary artery disease is result of ischemia and hypoxia of myocardial cells and endothelial cells of coronary artery. Stress, inflammatory reaction and lipidoese of ECs are the major mechanism of coronary artery disease. Another, angiogenesis take part in tumorigenesis, invasion and metabasis. In short, ECs is very important for many diseases. As we known, genes is same important in the process of diseases. What will induce the difference expression of genes of ECs in the process of diseases? What is its effect? To date, there is not overall analysis about gene expression of ECs. In the same time, this is significant for gene therapy.
Object:
To analysis all the gene expression profile of ECs through GeneChip of Affymetrix and get the important signals pathway, key genes and molecular function.
Methods:
(1)Gene expression profile of human umbilical vascular endothelial cells (HUVEC) is analyzed by the Affymetrix GeneChip HG-U133A plus2.0 and confirms the results of GeneChip with Real-time PCR.
(2)To mine the data by the bioinformatics software of internet. For example, Microsoft Excel, Milano, Go, PANTHER, Genomatix etal.
Results:
内皮细胞在人类许多重大疾病发生过程中处于重要的地位，例如冠心病、肿瘤等。冠心病是由于冠脉血管病变导致心肌细胞缺血、缺氧，使该支冠脉血管(包括微循环)及所支配区域的心肌细胞凋亡、坏死，其中内皮细胞的的应激、炎症反应、脂质沉积等病理生理过程是冠心病发生的重要机制。在肿瘤尤其是实体瘤的发生过程中，血管新生是肿瘤得以存活的重要因素，而血管新生的最主要过程即为内皮细胞的生长，另外肿瘤的侵袭、转移等过程，也有赖于内皮细胞的参与。因此可以看出，内皮细胞在这些关乎人民健康的重大疾病的发生过程中占有重要的地位。
基因在疾病的发生发展过程中处于重要的地位已成为科学界的共识。基因表达的改变在内皮细胞病变的过程中处于什么地位?发挥什么作用?——长期以来就是医学领域中研究的重点和热点。当前，对于内皮细胞基因表达方面的研究较多，但是尚未见将内皮细胞全部基因表达谱进行全部分析的报道。若能将内皮细胞的所有表达基因进行分析，汇总和分类，分析这些表达基因所参与的病理生理功能及在内皮细胞中所发挥的作用，那将对基因治疗运用于内皮细胞具有重要的意义。
目的：
利用当前基因芯片高通量技术，全面分析人脐静脉内皮细胞的基因表达谱，对所有表达的基因进行生物信息学分析，汇总整理出与内皮细胞相关的重要的基因、分子功能和信号通路。
方法：
(1) 应用人类HG-U133A plus 2.0芯片对人脐静脉内皮细胞的基因表达谱进行全面分析，进而就用荧光定量PCR方法对结果进行验证。
(2) 应用Microsoft Excel、Milano、Go、PANTHER以及Genomatix等软件和网站对基因表达谱进行分析、汇总及数据挖掘。
Introduction
Endothelial cells (ECs) are very important for many diseases, for example, coronary artery disease, and tumor. Coronary artery disease is result of ischemia and hypoxia of myocardial cells and endothelial cells of coronary artery. Stress, inflammatory reaction and lipidoese of ECs are the major mechanism of coronary artery disease. Another, angiogenesis take part in tumorigenesis, invasion and metabasis. In short, ECs is very important for many diseases. As we known, genes is same important in the process of diseases. What will induce the difference expression of genes of ECs in the process of diseases? What is its effect? To date, there is not overall analysis about gene expression of ECs. In the same time, this is significant for gene therapy.
Object:
To analysis all the gene expression profile of ECs through GeneChip of Affymetrix and get the important signals pathway, key genes and molecular function.
Methods:
(1)Gene expression profile of human umbilical vascular endothelial cells (HUVEC) is analyzed by the Affymetrix GeneChip HG-U133A plus2.0 and confirms the results of GeneChip with Real-time PCR.
(2)To mine the data by the bioinformatics software of internet. For example, Microsoft Excel, Milano, Go, PANTHER, Genomatix etal.
Results:
引文
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[3] Paris S, Denis H, Delaive E, et al. Up-regulation of 94-kDa glucose-regulated protein by hypoxia-inducible factor-1 in human endothelial cells in response to hypoxia. FEBS Lett, 2005.579(1): 105-14.
[4] Shi Y H, Wang Y X, Bingle L, et al. In vitro study of HIF-1 activation and VEGF release by bFGF in the T47D breast cancer cell line under normoxic conditions: involvement of PI-3K/Akt and MEK1/ERK pathways. J Pathol, 2005. 205(4): 530-6.
[5] Friedmann T. Human gene therapy--an immature genie, but certainly out of the bottle. Nat Med, 1996. 2(2): 144-7.
[6] Coutelle C, Themis M, Waddington S N, et al. Gene therapy progress and prospects: fetal gene therapy—first proofs of concept—some adverse effects. Gene Ther, 2005. 12(22): 1601-7.
[7] Seth P. Vector-mediated cancer gene therapy: an overview. Cancer Biol Ther, 2005. 4(5): 512-7.
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