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: 8332876
• 负责人: C. Michael DiPersio
• 金额: $ 23.04万
• 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 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8332876
• 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; Health; 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; Publishing; Regulation; Regulatory Element; Research; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site-Directed Mutagenesis; Specimen; Squamous Cell; Squamous cell carcinoma; Staging; System; 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 progression; 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.

描述（由申请人提供）：整合素a3b1是一种细胞外基质受体，在许多恶性肿瘤中表达，并已被证明可以调节与上皮间质转化（EMT）相关的细胞表型，例如细胞增殖、存活和侵袭。基质金属蛋白酶 MMP-9 的表达也与恶性肿瘤生长有关，可促进肿瘤血管生成和细胞侵袭。 a3b1 通过转录后 mRNA 稳定性在永生化角质形成细胞（MK 细胞）中诱导 MMP-9，这种调节是在细胞永生化过程中获得的。然而，a3b1 调节 MMP-9 mRNA 稳定性的机制和信号通路及其在肿瘤生长和进展中的作用尚不清楚。拟议研究的目标是通过利用一组表达 a3b1（即 a3 野生型）和 a3b1 缺陷（即 a3-null）的 MK 变体来回答这些问题，这些变体共同代表不同的 EMT 阶段：（1）从新生儿表皮中分离出永生化角质形成细胞； (2) 永生化 MK 细胞带有 p53 缺失突变； (2)转化的MK细胞另外表达致癌RasV12。 p53 缺失和致癌 Ras 激活是鳞状细胞癌和其他癌症中的常见突变。该模型的初步数据表明，a3b1 和 MMP-9 是 MK 细胞体内肿瘤生长所必需的，并确定了 a3b1 可能控制 MMP-9 mRNA 稳定性的候选信号传导途径和机制。拟议的研究将测试以下假设：MMP-9 mRNA 表达受控制 mRNA 稳定性的特定 a3b1 介导的信号通路控制，并且这些机制控制体内肿瘤生长。将结合分子、遗传和生化方法来鉴定控制 a3b1 依赖性 mRNA 稳定性的 mRNA 调控元件，并阐明参与 a3b1 介导的 MMP-9 mRNA 稳定性的特定信号传导途径或整合素功能。体内肿瘤发生模型将用于研究 MMP-9 的 a3b1 依赖性调节对肿瘤生长的作用。最后，将通过测试几种人类癌细胞系中 MMP-9 的 a3b1 依赖性调节，并评估人类肿瘤样本中 a3b1 和 MMP-9 的表达来研究这些观察结果的人类相关性。