Cell Biology Core

A.目标说明 细胞生物学核心是一个新设施，将为所有四个项目提供服务。这个核心将提供 需要研究的几个产生脂蛋白和非脂蛋白的细胞系 在每个项目中都有描述。 脂蛋白产生细胞将被用于：1)研究组装和 分泌内源性载脂蛋白A-L和载脂蛋白B-新生脂蛋白颗粒；2) 确定几种apoA-L模拟肽对含apoA-L和apoB的新生脂蛋白颗粒的浓度、组成和性质的影响；3)用于转染研究，以评估apoB和潜在的apoA-L及其各自的颗粒组装和分泌的结构功能关系；4)利用miRNA技术沉默目的基因；5)建立表达C末端截短型人apoB的细胞或缺乏特异性基因，即脂转移蛋白的细胞的克隆稳定转化子。 非脂蛋白产生细胞系将用于：1)ABCA1的表达， Abcg1和ABCG4；2)建立了高表达ABCA1的克隆稳定细胞系， 3)观察载脂蛋白A-L模拟肽对内毒素诱导的内毒素诱导的影响 THP-1单核细胞与人脐静脉内皮细胞的黏附；4)胆固醇外流研究。

A. STATEMENT OF OBJECTIVES The Cell Biology Core is a new facility and will serve all four projects. This core will provide several lipoprotein-producing and non-lipoprotein producing cell lines required for studies described in each project. The lipoprotein producing cells will be used: 1) To study the mechanisms of assembly and secretion of endogenous apoA-l- and apoB-containing nascent lipoprotein particles; 2) To determine the effects of several apoA-l mimetic peptides on the concentration, composition, and properties of apoA-l- and apoB-containing nascent lipoprotein particles; 3) For transfection studies to assess the structure-function relationship of apoB, and potentially apoA-l, and their respective particle assembly and secretion; 4) To silence the gene of interest using miRNA technique; 5) To establish clonal stable transformants of cells that express C-terminally truncated forms of human apoB or cells that lack specific gene, i.e, lipid transfer proteins. The non-lipoprotein producing cell lines will be used: 1) For the expression of ABCA1, ABCG1, and ABCG4; 2) To established clonal statble cell lines overexpressing ABCA1, ABCG1, and ABCG4; 3) To evaluate the effects of apoA-l mimetic peptides on LPS-induced adhesion of THP-1 monocytes to HUVECs; 4) For cholesterol efflux studies.