NOVEL SERPIN INHIBITOR OF ORAL SQUAMOUS CARCINOMA

口腔鳞状细胞癌的新型丝氨酸蛋白酶抑制剂

• 批准号: 6855146
• 负责人: JOHN J SAUK
• 金额: $ 35.27万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6855146
• 关键词: SCID mouse; SDS polyacrylamide gel electrophoresis; cell membrane; cell motility; clinical research; collagen; endoplasmic reticulum; extracellular matrix; flow cytometry; glycoproteins; human tissue; neoplasm /cancer invasiveness; neoplastic cell; protease inhibitor; protein structure function; serine proteinases; site directed mutagenesis; squamous cell carcinoma; stress proteins

DESCRIPTION (provided by applicant): The hypothesis that this proposal seeks to test is that CBP2/Hsp47 constitutes a portion of the regulatory complexity that determines whether collagen XVIll (col 18) is produced and processed to function as a motogen or as an endostatin (ES). Moreover, that CBP2/Hsp47 and col 18 form an integral component of the angiostatic-angiogenic switching axis that ensures tumor growth and promotes invasion. This proposal seeks to prove that during normoxia col 18 with the aid of CBP2/Hsp47 is produced and secreted where it may itself or processed by matrix metalloproteinases to motogenic fragments promote tumor cell motility and invasion. However, at low microenvironmental pH resulting from hypoxia or necrosis, col 18 is processed by cathepsin L, unless inhibited by extracellular CBP2/Hsp47, to ES. Endostatin inhibits tumor cell motility and invasion until and proangiogenic signaling establishes a neovasculature that reestablishes normoxia. The proposed research will be accomplished through the completion of the following specific aims: 1) demonstrate that short form of collagen XVIII (lacking the frizzled domain) promote tumor cell motility and invasion using in vitro models. Then show that proteolytic fragments of collagen XVilI, particularly the trimerized NC1 domain and fragments produced following matrix metalloproteinase activity are likewise motogenic while the carboxyl terminus, endostatin domain, inhibits both motility and invasion; 2) determine whether CBP2/Hsp47 constitutes an important feature of the regulatory complexity that controls collagen XVill by over expressing or knocking out CBP2/Hsp47 in human oral squamous carcinoma tumor cell lines; 3) Determine the signaling pathways by which collagen XVII! promotes tumor cell invasion and the feedback mechanism(s) by which its' C-terminal fragment, endostatin, functions as an antagonist and 4 )provide the proof in practice that in solid human tumors CBP2/Hsp47 and the microcellular environment constitute important features of the regulatory complexity that control patterns of tumor cell growth and invasion by regulating the production and processing of collagen XVIII. These studies will aid in correlating the gene transcription profiles, and repertoire of proteins expressed and their activities with functional status of both normal and aberrant cells.

描述（由申请人提供）：本提案试图测试的假设是，CBP2/Hsp47 构成了决定胶原蛋白 XVIll（第 18 列）是否被生产和加工以充当运动原或内皮抑素 (ES) 的调节复杂性的一部分。此外，CBP2/Hsp47 和 col 18 形成血管抑制-血管生成转换轴的一个组成部分，确保肿瘤生长并促进侵袭。该提案试图证明，在常氧条件下，CBP2/Hsp47 的帮助下产生和分泌 col 18，其本身或被基质金属蛋白酶加工成运动片段，促进肿瘤细胞运动和侵袭。然而，在缺氧或坏死导致的低微环境 pH 值下，除非受到细胞外 CBP2/Hsp47 的抑制，否则 Col 18 会被组织蛋白酶 L 加工成 ES。内皮抑素抑制肿瘤细胞的运动和侵袭，直到促血管生成信号建立新血管系统，从而重建正常氧浓度。拟议的研究将通过完成以下具体目标来完成：1）使用体外模型证明短形式的胶原蛋白 XVIII（缺乏卷曲结构域）可促进肿瘤细胞的运动和侵袭。然后表明胶原蛋白 XVilI 的蛋白水解片段，特别是三聚化的 NC1 结构域和基质金属蛋白酶活性后产生的片段同样具有运动原性，而羧基末端、内皮抑素结构域则抑制运动和侵袭； 2)通过在人口腔鳞癌肿瘤细胞系中过度表达或敲除CBP2/Hsp47来确定CBP2/Hsp47是否构成控制胶原蛋白XVill的调控复杂性的重要特征； 3) 确定胶原蛋白 XVII! 的信号传导途径！促进肿瘤细胞侵袭及其反馈机制，通过其 C 端片段内皮抑素发挥拮抗剂的作用，4）在实践中证明，在人类实体肿瘤中，CBP2/Hsp47 和微细胞环境构成了通过调节胶原蛋白 XVIII 的产生和加工来控制肿瘤细胞生长和侵袭模式的复杂性调节的重要特征。这些研究将有助于将基因转录谱、表达的蛋白质及其活性与正常和异常细胞的功能状态相关联。