HEPATIC AND COLONIC SIALYLTRANSFERASE EXPRESSION

肝脏和结肠唾液酸转移酶表达

• 批准号: 3464421
• 负责人: M. Peter Lance
• 金额: $ 10.02万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-06-01 至 1995-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3464421
• 关键词: RNA; athymic mouse; carbohydrate biosynthesis; cell differentiation; colorectal neoplasms; complementary DNA; dexamethasone; electrophoresis; enzyme mechanism; eukaryote; gene expression; genetic translation; glycoproteins; glycosyltransferase; hamsters; hepatocellular carcinoma; linkage mapping; messenger RNA; metastasis; neoplasm /cancer genetics; nucleic acid hybridization; nucleic acid probes; oligosaccharides; ovary; plasmids; posttranslational modifications; retinoate; sialyltransferases; thyroid hormones; tissue /cell culture

Sequential modification of the sialic acid component of cell adhesion molecules and other glycoproteins is one of the mechanisms by which normal cell differentiation proceeds. The sialyl oligosaccharides of aberrant glycoproteins have also been implicated in the pathophysiology of various diseases, including metastatic colorectal cancer. The synthesis of each sialyl linkage is catalyzed by a specific sialyltransferase and fluctuations in the activities of these enzymes may therefore play a critical role in such fundamental biological processes as differentiation and metastatic behavior. Towards an eventual understanding of the mechanisms that coordinate expression of the different sialyltransferases in normal and diseased tissues, objectives of this application are: to express cDNAs for human sialyltransferases in eucaryotic cells and delineate the sialyl linkages that are catalyzed by the different encoded proteins; to perform a tissue survey of human sialyltransferase mRNAs; to compare sialyltransferase mRNA expression and enzyme activity in a panel of human cancer cell lines; and to investigate in the same panel of cell lines the regulatory effects of agents that are likely to influence levels of sialyltransferase expression. Chinese hamster ovary cells will be transfected with constructs of the sialyltransferase cDNAs in plasmid vectors. The activities of expressed sialyltransferases will be characterized by determination of their acceptor substrates and analysis of the oligosaccharides that are synthesized. The tissues survey will be conducted in human surgical specimens through Northern hybridization analysis and S1 protection assays. The panel of cancer cells will include dexamethasone, n-butyrate, retinoic acid and thyroid hormone. The results will form the basis for an understanding of the determinants of the activities of sialyltransferases and future studies of the biological functions of sialylation.

细胞的唾液酸成分的顺序修饰 粘附分子和其他糖蛋白是一种机制之一 哪种正常细胞分化进行。 siAllyl寡糖的寡糖 异常糖蛋白也与 各种疾病，包括转移性结直肠癌。 合成 每个siAlyl链接都被特定的siAllltransferase催化 因此，这些酶的活动的波动可能会发挥作用 在区分等基本生物过程中的关键作用 和转移行为。 最终了解 协调不同硝基转移酶表达的机制 在正常和患病的组织中，此应用的目标是： 在念珠菌细胞和 描述由不同编码的催化的siAlyl链接 蛋白质；对人乙酰转移酶mRNA进行组织调查；到 比较一群小组中的乙烯基转移酶mRNA表达和酶活性 人类癌细胞系；并在相同的细胞系中进行调查 可能影响的代理的调节作用 辅助转移酶的表达。 中国仓鼠卵巢细胞将是 用质粒中的乙酰转移酶CDNA的构建体转染 向量。 表达的siallyltransferases的活动将是 以确定其受体底物和分析的特征 合成的寡糖。 组织调查将是 通过北部杂交在人类手术标本中进行 分析和S1保护测定法。 癌细胞面板将包括 地塞米松，N-丁酸，视黄酸和甲状腺激素。 结果 将构成理解决定因素的基础 乙烯酸转移酶的活性和生物学的未来研究 溶解的功能。