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: 6777419
• 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/6777419
• 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）：本提案旨在验证的假设是CBP2/Hsp47构成了调节复杂性的一部分，该复杂性决定了胶原xvi （col 18）是作为一种运动原还是作为一种内皮抑制素（ES）产生和加工。此外，CBP2/Hsp47和col 18构成了血管静止-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。本研究旨在证明在常氧条件下，col18在CBP2/Hsp47的帮助下产生和分泌，其可能自身或被基质金属蛋白酶加工成促运动片段，促进肿瘤细胞的运动和侵袭。然而，在缺氧或坏死导致的低微环境pH下，除非受到细胞外CBP2/Hsp47的抑制，否则col18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一个新的血管系统，重建正常氧合。本研究将通过完成以下具体目标来完成：1)利用体外模型证明短形式的胶原XVIII（缺乏卷曲结构域）促进肿瘤细胞的运动和侵袭。然后表明胶原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性，而羧基端内皮抑素结构域抑制运动性和侵袭性；2)通过在人口腔鳞癌细胞系中过表达或敲除CBP2/Hsp47，确定CBP2/Hsp47是否构成控制xvi胶原调控复杂性的重要特征；3)确定胶原蛋白XVII！促进肿瘤细胞侵袭及其c端片段内皮抑素作为拮抗剂的反馈机制；4)在实践中证明，在实体人肿瘤中，CBP2/Hsp47和微细胞环境是通过调节胶原蛋白的产生和加工来控制肿瘤细胞生长和侵袭模式的调控复杂性的重要特征。这些研究将有助于将基因转录谱、表达的蛋白库及其活性与正常和异常细胞的功能状态联系起来。