Cell Biology Core

A.目标说明 细胞生物学核心是一个新的设施，将服务于所有四个项目。这个核心将提供 研究所需的几种产生脂蛋白和不产生脂蛋白的细胞系 在每个项目中描述。 将使用脂蛋白产生细胞：1）研究组装和 分泌内源性含有apoA-I和apoB的新生脂蛋白颗粒; 2） 确定几种apoA-1模拟肽对浓度、组成和浓度的影响。 含有apoA-I和apoB的新生脂蛋白颗粒的性质; 3）对于转染 评估apoB和潜在的apoA-I的结构-功能关系的研究，以及它们的 4）使用miRNA沉默目的基因 技术; 5）建立表达C-末端的细胞的克隆稳定转化体， 截短形式的人apoB或缺乏特定基因的细胞，即脂质转移蛋白。 将使用不产生脂蛋白的细胞系：1）对于ABCA 1的表达， ABCG 1和ABCG 4; 2）为了建立过表达ABCA 1的克隆稳定细胞系， ABCG 1和ABCG 4; 3）评价apoA-1模拟肽对LPS诱导的小鼠肝癌细胞凋亡的影响， THP-1单核细胞与HUVEC的粘附; 4）用于胆固醇流出研究。

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.