BIOLOGY OF ENDOTHELIAL CELL DIFFERENTIATION

血管生成，即形成新血管的过程，是 基本生理和病理过程的数量，如伤口 愈合、胎盘发育和肿瘤转移。内皮细胞， 它们构成了所有血管的内层，似乎扮演着 在这一过程中发挥作用。通过改变他们所处的条件 培养的人脐静脉内皮细胞(HUVECs)可以被诱导 在体外形成毛细血管样网络。使用减法/放大 本实验室建立的方法，在细胞中差异表达的mRNA 与作为单层生长的HUVEC相比，区分HUVEC将是 分离、聚合酶链式反应扩增并用于差异筛选 从PMA处理的人脐静脉内皮细胞和分化的人脐静脉内皮细胞构建文库。 将对独特的cDNA进行测序并对其进行预测 结构/功能。用不同细胞的RNA进行Northern杂交 线条和组织将决定克隆的组织分布。 差异表达的蛋白质的功能将是 通过添加反义寡聚体或抗体来阐明的 细胞培养及自发性地将克隆导入 转化的人内皮细胞系。杂交种 荧光素标记的正常组织抗体和放射性核糖核酸 切片和病理标本(实验诱导的肿瘤 大鼠慢性关节炎)将有助于阐明该小说的作用 健康和疾病中的蛋白质。

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.