Regulation of MMP-9 mRNA stability and tumor growth by alpha3 beta1 integrin

α3β1 整合素对 MMP-9 mRNA 稳定性和肿瘤生长的调节

• 批准号: 7474354
• 负责人: C. Michael DiPersio
• 金额: $ 32.58万
• 依托单位: ALBANY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-26 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7474354
• 关键词: Adhesions; Binding; Biochemical; CD29 Antigen; Carcinoma; Cell Proliferation; Cell model; Cells; Data; Development; Dominant-Negative Mutation; ECM receptor; Elements; Epidermis; Epithelial; Epithelial Cells; Extracellular Matrix; Family; Gelatinase B; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic; Goals; Guanosine Triphosphate Phosphohydrolases; Human; Indium; Integrins; Laminin; Link; MAP Kinase Gene; MAPK14 gene; Malignant - descriptor; Malignant Neoplasms; Matrix Metalloproteinases; Mediating; Messenger RNA; Modeling; Molecular; Molecular Genetics; Molecular Target; Mutation; Neonatal; Neoplasm Metastasis; Normal Cell; Nude Mice; Oncogenic; Pathway interactions; Phenotype; Polyadenylation; Public Health; Publishing; Regulation; Regulatory Element; Research; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site-Directed Mutagenesis; Specimen; Squamous Cell; Squamous cell carcinoma; Staging; System; TP53 gene; Testing; Transcript; Tumor Angiogenesis; Tumor Cell Invasion; Untranslated Regions; Variant; base; cancer cell; cancer therapy; cell growth; data modeling; epithelial to mesenchymal transition; in vivo; keratinocyte; mRNA Expression; mRNA Stability; neoplastic cell; novel; null mutation; receptor; research study; rho; small hairpin RNA; therapeutic target; tumor; tumor growth; tumor necrosis factor receptor superfamily, member 10b protein, mouse; tumorigenesis; tumorigenic

DESCRIPTION (provided by applicant): Integrin a3b1 is an extracellular matrix receptor that is expressed in many malignant tumors and has been shown to regulate cellular phenotypes associated with epithelial-to-mesenchymal transition (EMT), such as cell proliferation, survival, and invasion. Expression of matrix metalloproteinase MMP-9 is also linked to malignant tumor growth, where it promotes tumor angiogenesis and cell invasion. a3b1 induces MMP-9 in immortalized keratinocytes (MK cells) through post-transcriptional mRNA stability, and this regulation is acquired during cellular immortalization. However, the mechanisms and signaling pathways whereby a3b1 regulates MMP-9 mRNA stability and their roles in tumor growth and progression are unknown. The goal of the proposed research is to answer these questions by exploiting a panel of a3b1-expressing (i.e. a3 wild type) and a3b1- deficient (i.e., a3-null) MK variants that collectively represent different EMT stages: (1) non-immortalized keratinocytes are isolated from neonatal epidermis; (2) immortalized MK cells harbor a p53-null mutation; (2) transformed MK cells additionally express oncogenic RasV12. Loss of p53 and oncogenic Ras activation are common mutations in squamous cell and other carcinomas. Preliminary data from this model indicate that a3b1 and MMP-9 are required for in vivo tumor growth of MK cells, and identify candidate signaling pathways and mechanisms whereby a3b1 may control MMP-9 mRNA stability. The proposed studies will test the hypotheses that MMP-9 mRNA expression is controlled by specific a3b1-mediated signaling pathways that control mRNA stability, and that these mechanisms control tumor growth in vivo. A combination of molecular, genetic, and biochemical approaches will be used to identify mRNA regulatory elements that control a3b1- dependent mRNA stability, and to elucidate specific signaling pathways or integrin functions that are involved in a3b1-mediated MMP-9 mRNA stability. An in vivo tumorigenesis model will be used to investigate the role of a3b1-dependent regulation of MMP-9 for tumor growth. Finally, the human relevance of these observations will be investigated by testing for a3b1-dependent regulation of MMP-9 in several human carcinoma lines, and by assessing expression of a3b1 and MMP-9 in human tumor specimens. PUBLIC HEALTH RELEVANCE: A key to the development of anti-cancer therapies is the identification of molecular targets that are required for tumor growth, progression, and metastasis. The proposed studies will identify novel molecular pathways that are turned on in cancer cells to promote malignant tumor growth and metastasis. These pathways may be exploitable as therapeutic targets.

描述(申请人提供)：整合素a3b1是一种细胞外基质受体，在许多恶性肿瘤中表达，并已被证明调节与上皮向间充质转化(EMT)相关的细胞表型，如细胞增殖、存活和侵袭。基质金属蛋白酶-9的表达也与恶性肿瘤的生长有关，促进肿瘤血管生成和细胞侵袭。A3b1通过转录后稳定的mRNA诱导永生化角质形成细胞(MK细胞)中的基质金属蛋白酶-9，这种调节是在细胞永生化过程中获得的。然而，a3b1调节基质金属蛋白酶-9mRNA稳定性的机制和信号通路以及它们在肿瘤生长和进展中的作用尚不清楚。这项研究的目的是通过利用一组表达a3b1(即A3野生型)和a3b1缺陷(即a3b1缺失)的MK变体来回答这些问题，这些变体共同代表了不同的EMT阶段：(1)未永生化的角质形成细胞从新生儿表皮中分离出来；(2)永生化的MK细胞存在P53零突变；(2)转化的MK细胞额外表达癌基因RasV12。P53缺失和致癌RAS激活是鳞状细胞癌和其他癌组织中常见的突变。该模型的初步数据表明，A3b1和MMP9是MK细胞体内肿瘤生长所必需的，并确定了A3b1控制MMP9mRNA稳定性的候选信号通路和机制。这项拟议的研究将检验这样的假设，即基质金属蛋白酶-9mRNA的表达是由特定的a3b1介导的信号通路控制的，这些信号通路控制着mRNA的稳定性，并且这些机制控制着体内的肿瘤生长。结合分子、遗传和生物化学方法，将被用来确定控制依赖于3b1的信使核糖核酸稳定性的信使核糖核酸调节元件，并阐明涉及到信使通路或整合素功能的依赖于3b1的基质金属蛋白酶-9信使核糖核酸的稳定性。体内肿瘤发生模型将被用来研究依赖于3b1的基质金属蛋白酶-9对肿瘤生长的调节作用。最后，将通过在几个人类癌细胞系中检测依赖于3b1的基质金属蛋白酶-9的调节，以及通过评估人类肿瘤标本中a3b1和基质金属蛋白酶-9的表达来研究这些观察结果与人类的相关性。公共卫生相关性：抗癌治疗发展的一个关键是确定肿瘤生长、进展和转移所需的分子靶点。这项拟议的研究将确定在癌细胞中启动的促进恶性肿瘤生长和转移的新的分子途径。这些途径可能被开发为治疗靶点。