Chemoresistance and Motility: Role of Mutant p53 and NF-kB2 in Cancer

化疗耐药性和运动性：突变体 p53 和 NF-kB2 在癌症中的作用

• 批准号: 7898567
• 负责人: Sumitra Deb
• 金额: $ 24.82万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7898567
• 关键词: Affect; Biological Assay; Breast Cancer Cell; Breast Cancer Treatment; Cancer cell line; Carcinoma; Cell Adhesion; Cell Culture Techniques; Cells; Clinic; Cultured Cells; Data; Development; Doctor of Philosophy; Drug usage; Etoposide; Event; Future; Gene Expression Profiling; Gene Targeting; Genes; Genetic Transcription; Goals; H1299; Human; IL8 gene; Immunoprecipitation; Laboratories; Lead; Lung; Lung Neoplasms; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of lung; Modeling; Molecular; Mutation; NF-kappa B; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Nude Mouse Assay; Oncogenic; Pathway interactions; Patients; Phenotype; Play; Property; Protein p53; Proteins; RNA; RNA Interference; Research; Resistance; Role; Sampling; Specimen; TP53 gene; Testing; Transactivation; Transcriptional Activation; Transfection; Tumor Suppressor Genes; Tumor Suppressor Proteins; Tumor-Derived; Tumorigenicity; Up-Regulation; angiogenesis; base; c-myc Genes; cancer cell; cell growth; cell motility; chemokine; chemotherapeutic agent; chromatin immunoprecipitation; cyclin B2; cyclin D2; gain of function; in vivo; inhibitor/antagonist; lung Carcinoma; malignant breast neoplasm; mutant; neovascularization; novel; overexpression; pressure; prevent; promoter; protein protein interaction; public health relevance; transcription factor; tumor; tumor progression; tumorigenesis; tumorigenic; vector

DESCRIPTION (provided by applicant): Mutation in the p53 tumor suppressor gene is a common event in human cancer. In the majority of human carcinomas with p53 mutations the mutant protein is over-expressed suggesting the existence of a selective advantage to maintain expression. The long-term goal of our research is to understand how p53 mutations lead to oncogenesis. The short-term objective is to test the following hypothesis: Expression of p53 mutants in human cells deregulates pathways controlled by the NF-kB2, a property critically important for chemosensitivity and tumor progression. The above-mentioned hypothesis is based on the following observations: Compared to vector transfected cells, H1299 p53-null human lung carcinoma cells expressing mutant p53 showed chemo-resistance when treated with common chemotherapeutic agents; however, cells expressing transactivation deficient mutants lose this function significantly, suggesting that transactivation by mutant p53 is crucial for chemo-resistance. A number of lung and breast cancer cell line with mutant p53 show chemo-resistance that is dependent on the level of p53 as the chemo-resistance decreases when the p53 level is lowered. p53 mutants induced expression of a number of genes involved in cell growth, survival, invasion, metastasis and angiogenesis. NF-kB2 was among this group. Introduction of short interfering RNA specific for NF-:B2 made these cells lose chemo-resistance. A preliminary screen of human lung cancer specimens shows co-existing p53 mutation and over-expression of NF-kB2 suggesting that, in the clinic, there is a subset of patients with p53 mutation and NF-kB2 over-expression. Our data also indicate that mutant p53 expression enhances cell adhesion, motility, tumorigenicity and metastatic phenotype. The following are the specific aims: 1. (a) To determine whether mutant p53 over-expression leads to higher levels of NF-kB2 in lung cancer. (b) To determine whether NF-kB2 is involved in reducing chemosensitivity in cancer cells expressing mutant p53. 2. To determine whether NF-:B2 enhances motility of cells expressing mutant p53. 3. To determine the mechanism of NF-kB2 up-regulation by mutant p53. 4. To identify factors interacting specifically with GOF p53 mutants utilizing mass spectrometric analysis. The proposed research will investigate the relationship between p53 mutants commonly found in cancer and the NF-:B2 pathway. In future, tumors with p53 mutations can be targeted at NF-:B2, mutant p53 and many of their potential target genes. Chemoresistance and Motility: Role of Mutant p53 and NF-:B2 in Cancer. PUBLIC HEALTH RELEVANCE: This application is based on a novel observation that tumor-derived p53 mutants up-regulate NF-kB2 expression in cell culture. Preliminary examination of human tumors also indicates that lung tumors with p53 mutation have a higher NF-:B2 level. We also find that mutant p53 expression induces higher motility and chemo-resistance in cells expressing them. We propose to determine the molecular mechanism of this up- regulation and find out how that is related to chemo-resistance and higher motility of cells expressing mutant p53. Results obtained from this study should lead to identification of a subset of lung tumors co- expressing NFkB2 and mutant p53 that could be targeted by inhibitors of NF-kB pathway to prevent tumor progression.

描述（由申请人提供）：p53肿瘤抑制基因中的突变是人类癌症中的常见事件。在大多数具有p53突变的人类癌中，突变蛋白过表达表明存在选择性优势以维持表达。我们研究的长期目标是了解p53突变如何导致肿瘤发生。短期目标是检验以下假设：人类细胞中p53突变体的表达取消了由NF-KB2控制的途径，NF-KB2对化学敏感性和肿瘤进展至关重要。上述假设基于以下观察结果：与载体转染的细胞相比，表达突变体p53的H1299 p53-null人类肺癌细胞在用常见的化学治疗剂治疗时表现出化学抗性；然而，表达反式激活缺陷突变体的细胞大大损失了这一功能，这表明突变体p53的反式激活对于化学抗性至关重要。许多带有突变体p53的肺和乳腺癌细胞系显示化学抗性，这取决于p53的水平，因为当p53水平降低时，化学抗性降低。 p53突变体诱导许多与细胞生长，生存，侵袭，转移和血管生成有关的基因的表达。 NF-KB2是该组之一。引入简短干扰RNA的NF-：B2使这些细胞失去了化学抗性。人肺癌标本的初步筛查显示，NF-KB2的共存P53突变和过表达表明，在诊所中，有一部分患有p53突变和NF-KB2过表达的患者。我们的数据还表明，突变体p53表达增强了细胞粘附，运动性，肿瘤性和转移表型。以下是具体目的：1。（a）确定突变体p53过表达是否导致肺癌中NF-KB2的水平较高。 （b）确定NF-KB2是否参与降低表达突变体p53的癌细胞的化学敏感性。 2。确定NF-：B2是否增强了表达突变体p53的细胞的运动。 3。确定突变体p53上调NF-KB2上调的机理。 4。确定使用质谱分析的GOF p53突变体特别相互作用的因素。拟议的研究将研究癌症中常见的p53突变体与NF-：B2途径之间的关系。将来，具有p53突变的肿瘤可以针对NF-：B2，突变体P53及其许多潜在靶基因。化学耐药性和运动性：突变体p53和NF-：B2在癌症中的作用。公共卫生相关性：该应用是基于一个新的观察结果，即肿瘤衍生的p53突变体在细胞培养中上调NF-KB2表达。对人肿瘤的初步检查还表明，具有p53突变的肺肿瘤具有较高的NF-：B2水平。我们还发现，突变体p53表达在表达它们的细胞中诱导更高的运动性和化学抗性。我们建议确定这种上调节的分子机制，并找出与表达突变体p53细胞的化学抗性和更高运动性有关的。从这项研究中获得的结果应导致鉴定共同表达NFKB2和突变体p53的子集，这些肺部肿瘤可以由NF-KB途径的抑制剂靶向以防止肿瘤进展。