Anlaysis of SPARC in cell invasion in C. elegans

SPARC 在秀丽隐杆线虫细胞侵袭中的分析

• 批准号: 7274628
• 负责人: GINGER R MILEY
• 金额: $ 5.13万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-28 至 2009-11-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7274628
• 关键词: Anlaysis; SPARC; cell; invasion; C.

DESCRIPTION (provided by applicant): Cell invasion across basement membranes is a driving force behind cell dispersal and organ formation during normal development, but also is used by metastatic cancer cells. The mechanisms used by cells to invade remain poorly understood. Therefore, elucidating the mechanisms that mediate cell invasion is medically significant. Cell invasion is a multi-step process that involves: 1) attachment of the invading cell to the basement membrane, 2) removal of the basement membrane, and 3) migration through the basement membrane. SPARC is an extracellular matrix (ECM) protein that has multiple functions in development and is overexpressed in a number of metastatic cancers. We have discovered that SPARC promotes anchor cell (AC) invasion across basement membranes in Caenorhabditis elegans. We hypothesize that SPARC.may function to alter the basement membrane or other ECM proteins to promote AC invasion. This suggests a new mechanism by which SPARC facilitates metastasis during cancer progression by fostering invasion across basement membranes in cells that would normally be incapable of invasion. C. elegans is a simple model system allowing for a detailed, in vivo analysis to study the ability of SPARC to promote cell invasion. This proposal will investigate the role of SPARC by 1) identifying the temporal and functional requirements of SPARC in fostering AC invasion, 2) determining if SPARC promotes invasion by altering the basement membrane and ECM proteins, and 3) testing whether SPARC functions in the AC and regulates the expression of AC target genes. A combination of approaches will be used including RNA interference, AC- specific promoters, time-lapse analysis, fluorescence-labeling experiments, and immunohistochemistry. This will allow for a deeper understanding into the function of SPARC in cell invasion and basement membranes, and will provide the candidate with new training in ECM biology, advanced microscopy, and cell biology. Relevance to public health: Not only are cell invasion events needed for proper development, but cancerous cells exploit these same mechanisms in order to metastasize although these mechanisms are poorly understood. The ECM protein SPARC is overexpressed in numerous metastatic cancers and our preliminary research suggests SPARC fosters cell invasion in the nematode worm Caenorhabditis elegans, which shares molecular and cellular basement membrane aspects of invasive metastatic cancer cells. The aim of this work is to elucidate the mechanism(s) underlying the ability of SPARC to promote cell invasion, which will likely lead to the development of more effective strategies to treat cancer.

描述（由申请人提供）：跨基底膜的细胞侵袭是正常发育过程中细胞扩散和器官形成背后的驱动力，但也被转移性癌细胞使用。细胞用来侵袭的机制仍然了解不足。因此，阐明介导细胞侵袭的机制具有医学意义。细胞侵袭是一个多步骤的过程，涉及：1）入侵细胞在基底膜上的附着，2）去除基底膜，3）3）通过基底膜迁移。 SPARC是一种细胞外基质（ECM）蛋白，在发育中具有多个功能，并且在许多转移性癌症中过表达。我们发现，SPARC促进了秀丽隐杆线虫的锚固细胞（AC）侵袭。我们假设SPARC.MAY功能可以改变基底膜或其他ECM蛋白来促进AC侵袭。这表明了一种新的机制，通过促进细胞中跨基底膜的侵袭通常无法入侵，SPARC通过促进癌症进展过程中的转移。秀丽隐杆线虫是一个简单的模型系统，允许进行详细的体内分析，以研究SPARC促进细胞侵袭的能力。该提案将通过1）确定SPARC在促进AC侵袭中的时间和功能需求来研究SPARC的作用，2）确定SPARC是否通过改变基础膜和ECM蛋白来促进侵袭，以及3）测试SPARC是否在AC中的功能是否在AC中功能并调节AC靶基因的表达。将使用多种方法，包括RNA干扰，特定的启动子，延时分析，荧光标记实验和免疫组织化学。这将使对SPARC在细胞侵袭和基底膜中的功能有更深入的了解，并将为候选人提供ECM生物学，晚期显微镜和细胞生物学的新培训。与公共卫生的相关性：不仅适当发育需要细胞侵袭事件，而且癌细胞利用这些相同的机制来转移，尽管这些机制知之甚少。 ECM蛋白SPARC在许多转移性癌症中过表达，我们的初步研究表明，SPARC促进了线虫蠕虫Caenorhabditis elegans中的细胞侵袭，该细胞虫具有分子和细胞基底膜的膜状膜膜的侵袭性转移性癌细胞。这项工作的目的是阐明SPARC促进细胞侵袭能力的基本机制，这可能会导致发展更有效的治疗癌症的策略。