CANCER AND HYPERCALCEMIA

该方案旨在研究生物合成，分泌，细胞- 甲状旁腺相关的特异性加工和基因表达 蛋白（PTHrP）与恶性肿瘤伴高钙血症相关， 免疫化学和核酸杂交方法， 用于PTHrP的临床诊断、分类和管理- 产生肿瘤。 PTHrP是一种141个残基的蛋白质， 肿瘤通常与高钙血症有关。 前13种氨基酸 在该蛋白质中，8个与人PTH的氨基末端同源， PTH的生物活性片段。 PTHrP的氨基末端片段， PTHrP 1 -34，与PTH受体反应，产生大部分的生物活性。 天然PTH的影响，包括高钙血症。 我们开发了 实验程序，研究细胞和分子生物学， PTHrP。 我们已经开发了几种细胞系， PTHrP mRNAS和PTHrP本身。 我们有特异性抗体 在组织和生物体液中鉴定PYHrP， 探针来研究其基因表达。 在免疫组织学研究中， 能够证明PTHrP在各种组织，包括鳞状细胞癌， 细胞肺癌和小细胞肺癌。 我们建议研究 在这些细胞系中PTHrP的生物合成和分泌， 放射免疫测定和相关免疫化学方法。 的 放射免疫测定程序将另外在两个中心进行 格式，以测量PTHrP的定义种类。这些试验研究 将与规模调整程序一起进行，例如 蛋白质分析和脉冲追踪实验。 的 通过北方杂交评价PTHrP mRNA的调节。 在 除了这些人体组织，我们还开发了动物模型， PTHrP研究 这些实验工具将允许基本的监管 我们提出的研究。 我们还建议研究 PTHrP作为内分泌癌的组织和血清标志物 开发了基于多表位识别的免疫化学程序。 这些研究将有助于阐明PTHrP在分子， 肺肿瘤的细胞和临床生物学 特色

This proposal is designed to study the biosynthesis, secretion, cell- specific processing, and gene expression of the parathyroid hormone-related protein (PTHrP) associated with the hypercalcemia of malignancy with immunochemical and nucleic acid hybridization procedures that can also be used in the clinical diagnosis, classification, and management of PTHrP- producing tumors. PTHrP is a 141 residue protein that is produced by tumors commonly associated with hypercalcemia. Of the first 13 amino acids of this protein, 8 are homologous to the amino terminus of human PTH, the biologically active fragment of PTH. The amino-terminal fragment of PTHrP, PTHrP1-34, reacts with the PTH receptor and produces most of the biological effects of native PTH, including hypercalcemia. We have developed the experimental procedures to study the cellular and molecular biology of PTHrP. We have developed several cell lines that produce multiple species of PTHrP mRNAS and PTHrP, itself. We have specific antibodies that identify PYHrP in tissues and biological fluids and we have nucleotide probes to study its gene expression. In immunohistology studies, we have been able to demonstrate PTHrP in a variety of tissues, including squamous cell lung cancer and also small-cell lung cancer. We propose to study the biosynthesis and secretion of PTHrP in these cell lines using radioimmunoassay and related immunochemical procedures. The radioimmunoassay procedures will additionally be performed in a two-site format in order to measure defined species of PTHrP. These assay studies will be conducted in conjunction with sizing procedures such as chromatography, Western Analysis, and pulse-chase experiments. The regulation of PTHrP mRNA will be evaluated by Northern hybridization. In addition to these human tissues, we have developed animal models for the study of PTHrP. These experimental tools will permit the basic regulatory studies that we propose. We also propose to study the clinical role of PTHrP as a tissue and serum marker for endocrine cancers with newly developed immunochemical procedures based on multiple epitope recognition. These studies should help elucidate the role of PTHrP in the molecular, cellular, and clinical biology of lung tumors with endocrine characteristics.