Role of Fetuin-A in Tumor Cell Growth

胎球蛋白-A 在肿瘤细胞生长中的作用

基本信息

批准号：
7942250
负责人：
JOSIAH OCHIENG
金额：
$ 16.29万
依托单位：
MEHARRY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7942250
关键词：
1-Phosphatidylinositol 3-Kinase A Mouse Acids Address Affect Amino Acid Sequence Animals Annexins Area Back Binding Biological Brain Breast Carcinoma Calcium Cell Communication Cell Surface Receptors Cell surface Cells Cellular biology Code Complex Confocal Microscopy Control Animal Culture Media Data Drug Design Eligibility Determination Esophageal Galectin 3 Genes Glycoproteins Goals Growth Growth Factor In Vitro Ions Knockout Mice LNCaP Lectin Lewis Lung Carcinoma Liver Lung Malignant Epithelial Cell Malignant neoplasm of prostate Mammary Neoplasms Mediating Names National Institute of General Medical Sciences Neoplasm Metastasis PC3 cell line Pancreas Pathway interactions Pharmaceutical Preparations Phosphorylation Phosphotransferases Reading Resistance Role Serum Serum Proteins Sialic Acids Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Molecule Skin Specificity Surface Techniques Testing Transferrin Transforming Growth Factor beta Work alpha-Fetoproteins base cancer cell cancer type cell growth frontier growth promoting activity in vivo inhibitor/antagonist innovation knock-down neoplastic cell receptor sugar treatment strategy tumor tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): Our long-term objectives are to elucidate the mechanisms by which fetuin-A, a serum glycoprotein synthesized by the liver, mediates and regulates the in vitro and in vivo growth of tumor cells. The central hypothesis is that fetuin-A derived from the serum or synthesized by the tumor cells interacts with its putative cell surface receptors, triggering PI3 kinase/Akt signaling that drives the growth of the cells. The hypothesis will be investigated through 4 specific aims: (1) To define the cell surface receptors for fetuin-A in tumor cells responsible for growth signaling; (2) To determine other signaling pathways which are modulated in tumor cells by the annexin/fetuin-A interaction; (3) To elucidate the role of ectopically synthesized fetuin-A in tumor cell growth; and (4) To define the specificity of serum derived fetuin-A in tumor growth promotion. In aim 1, we will extend our studies that were done on a limited scale using LLC cells to LNCaP prostate cancer cell line transfected with annexin-2-GFP. The co-localization and interaction between annexin-2 and fetuin-A in these cells will be studied using confocal microscopy. It is expected that annexins will emerge as the cell surface receptors for fetuin-A responsible for growth signaling. In aim 2, Kinexus and Affymetrix gene arrays will be used to screen for other signaling molecules whose expression is affected by the fetuin-A/annexin interaction. A number of signaling pathways cross-talk with the PI3 kinase/Akt pathway that is predominantly activated by fetuin-A. In aim 3, fetuin-A gene will be added back to cells that do not express it to determine if this is sufficient to allow these cells to grow under serum-free conditions and in fetuin-A null mice. Likewise, fetuin-A expression in LLC cells will be knocked down by sh-RNA approach and their growth under serum-free conditions re-evaluated. In aim 4, the serum from fetuin-A wild-type animals will be used to supplement growth medium to determine its growth promoting abilities in a variety of tumor cells vis-a-vis growth medium supplemented with serum from fetuin-A null mice. In this aim, we will determine whether the unique ability of fetuin-A to promote cell growth is also dictated by its amino acid sequence and not merely the presence of sialic acid residues. This proposal seeks to demonstrate that fetuin-A is an important fuel for the in vivo growth of tumor cells and is not an innocent bystander. The full understanding of this pathway has broad implications in the management and treatment strategies for majority of cancer types.

描述（由申请人提供）：我们的长期目标是阐明肝脏合成的血清糖蛋白（血清糖蛋白）合成的机制，可介导并调节肿瘤细胞的体外和体内生长。中心假设是源自血清的fetuin-a或由肿瘤细胞合成的fetuin-a与其推定的细胞表面受体相互作用，从而触发了驱动细胞生长的PI3激酶/AKT信号传导。该假设将通过4个特定目的进行研究：（1）定义负责生长信号传导的肿瘤细胞中fetuin-A的细胞表面受体；（2）确定通过膜联蛋白/fetuin-A相互作用在肿瘤细胞中调节的其他信号通路；（3）阐明异位合成fetuin-A在肿瘤细胞生长中的作用；（4）定义肿瘤生长促进中血清衍生的fetuin-a的特异性。在AIM 1中，我们将扩展使用LLC细胞在有限尺度上进行的研究，以将Annexin-2-GFP转染的LNCAP前列腺癌细胞系进行。这些细胞中的膜联蛋白-2和Fetuin-A之间的共定位和相互作用将使用共聚焦显微镜研究。可以预期，膜联蛋白将成为fetuin-a负责生长信号的细胞表面受体。在AIM 2中，Kinexus和Affymetrix基因阵列将用于筛选其他信号分子，其表达受fetuin-a/Annexin相互作用的影响。许多信号通路与PI3激酶/AKT途径交叉，主要由fetuin-a激活。在AIM 3中，将添加FETUIN-A基因回到未表达其表达的细胞中，以确定这是否足以允许这些细胞在无血清条件下和Fetuin-A-A无效小鼠中生长。同样，LLC细胞中的Fetuin-A表达将被SH-RNA方法击倒，并在无血清条件下重新评估其生长。在AIM 4中，将使用fetuin-A野生型动物的血清来补充生长培养基，以确定其在多种肿瘤细胞中促进生长能力，该肿瘤细胞与补充了fetuin-a无效小鼠的血清生长培养基。在此目标中，我们将确定fetuin-A促进细胞生长的独特能力是否还取决于其氨基酸序列，而不仅仅是唾液酸残基的存在。该提议试图证明fetuin-a是肿瘤细胞体内生长的重要燃料，并且不是无辜的旁观者。对这一途径的全面理解对大多数癌症类型的管理和治疗策略具有广泛的影响。