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Cell Biology Core

细胞生物学核心

基本信息

批准号：
8242750
负责人：
NASSRIN DASHTI
金额：
$ 23.96万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-01 至 2013-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8242750
关键词：
Adhesions Air American Type Culture Collection Anabolism Animals Apolipoproteins Apolipoproteins A Apolipoproteins B Atherosclerosis Benzamidines Biochemical Biological Assay Caco-2 Cells Carbon Dioxide Catabolism Cell Line Cells Cellular biology Chloramphenicol Cholesterol Conditioned Culture Media Cytolysis Edetic Acid Experimental Models Gene Silencing Genes Glycerol Hepatic Hepatoblastoma High Density Lipoprotein Cholesterol High Density Lipoproteins Human Human Cell Line In Vitro Incubated Inorganic Sulfates Intestines Label Lipids Lipoproteins Liver Liver parenchyma Low-Density Lipoproteins Maintenance Measurement Mediating Metabolism Methionine MicroRNAs Nutritional Penicillin G Peptides Phenylmethylsulfonyl Fluoride Phosphate Buffer Phospholipids Plasma Process Production Property Proteins Regulation Research Design Saline Sampling Serum Staging Streptomycin Structure-Activity Relationship Study models Techniques Testing Time Transfection Unspecified or Sulfate Ion Sulfates Very low density lipoprotein fetal bovine serum hexanoic acid hormone regulation in vivo interest lipid transfer protein mimetics monocyte monolayer overexpression particle peptide A peptide L planetary Atmosphere prevent research study tissue culture

项目摘要

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.

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模拟肽对含有apoA-1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-1的结构-功能关系，以及它们各自的颗粒组装和分泌; 4）使用miRNA技术沉默感兴趣的基因; 5）建立表达C-末端截短形式的人apoB的细胞或缺乏特异性基因即脂质转移蛋白的细胞的克隆稳定转化体。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。