THE BIOLOGY OF ENDOTHELIAL CELL DIFFERENTIATION

血管生成，即形成新血管的过程，是血管生成的核心。 许多基本的生理和病理过程，如创伤 愈合、胎盘发育和肿瘤转移。 内皮细胞， 形成了所有血管的内层， 在这个过程中的作用。 通过改变它们所处的环境 可以诱导生长的人脐静脉内皮细胞（HUVEC）， 在体外形成毛细血管样网络。 使用减法/放大 在这个实验室开发的方法，mRNA差异表达， 与作为单层生长的HUVEC相比，分化HUVEC将是 分离，通过PCR扩增，并用于差异筛选cDNA 从PMA处理的HUVEC和分化的HUVEC构建的文库。 将对独特的cDNA进行测序，并对其进行预测。 结构/功能。 用来自不同细胞的RNA进行北方印迹 品系和组织将决定克隆的组织分布。 将对差异表达蛋白的功能进行初步探讨 通过添加反义寡聚体或抗体来阐明， 细胞培养和转染克隆到自发的 转化的人内皮细胞系。 杂交 荧光素标记的抗体和放射性标记的RNA对正常组织 切片和病理标本（肿瘤，实验诱导 慢性关节炎大鼠）将有助于阐明的作用， 蛋白质在健康和疾病中的作用

Angiogenesis, the process of forming new blood vessels, is central to a number of essential physiologic and pathologic prosesses, such as wound healing, placental development and tumor metastasis. Endothelial cells, which form the lining of all blood vessels, appear to play a central role in this process. By altering the conditions under which they are grown, human umbilical vein endothelial cells (HUVECs) can be induced to form capillarylike networks in vitro. Using a subtraction/amplification method developed in this lab, mRNA differentially expressed in differentiating HUVECs compared to HUVECs growing as a monolayer will be isolated, amplified by PCR, and used to differentially screen cDNA libraries constructed from PMA-treated HUVECs and differentiated HUVECS. Unique cDNAs will be sequenced and predictions made concerning their structure/function. Northern blots performed with RNA from various cell lines and tissues will determine the tissue distribution of the clones. The functions of the differentially expressed proteins will be elucidated by the addiiion of antisense oligomers or antibodies added to cell cultures and transfection of the clones into a spontaneously transformed human endothelial cell line. Hybridization of fluorescein-labeled antibodies and radiolabed RNA to normal tissue sections and pathologic specimens (tumors, experimentally-induced chronic arthritis in rats) will help elucidate the role of the novel proteins in health and disease.