CHARACTERIZATION OF DIFFERENTIAL GENE EXPRESSION IN RESISTANT CELLS

耐药细胞中差异基因表达的特征

• 批准号: 3808200
• 负责人: ANATOLY DRITSCHILO
• 金额: --
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3808200
• 关键词: Escherichia coli; RNA; cellular pathology; electrophoresis; gene expression; genetic library; head /neck neoplasm; high performance liquid chromatography; molecular cloning; molecular oncology; neoplasm /cancer diagnosis; neoplasm /cancer genetics; neoplastic cell; nucleic acid hybridization; oncogenes; oncoproteins; plasmids; polymerase chain reaction; protein purification; radiation genetics; radiation sensitivity; squamous cell carcinoma; statistics /biometry; tissue /cell culture; transfection

Cell lines from cancer patients who undergo radiation therapy have been correlated to clinical outcomes and to in vitro radiation responses. That some cells of similar histologies, particularly from patients whose treatments have failed to locally control their tumors, show in vitro radiation survival parameters of a resistant type, while others are of a sensitive type, suggests a genetic basis for radiation sensitivity. The overall objective of this project is to identify genes that may be associated with the radiation resistance of human tumors. Using the techniques of protein analysis (2-D gel electrophoresis and microsequencing) differential screening/subtraction cDNA library analysis, and vector mediated gene transfer, a multiple approach strategy is planned in the first AIM to identify and isolate genes associated with the radiation-resistant phenotype. We will test the hypothesis that differential gene expressions in cells are responsible for the radiation sensitive or the radiation resistant phenotypes of tumor cells. In the second AIM of this project, two strategies to identify genes associated with resistance will be used to modify the radiation responses of sensitive tumor cells using vector mediated gene transfer. The first will use an episomally maintained vector strategy containing cDNA from radiation resistant cells and the second will use a cosmid strategy for gene transfer using larger fragments of DNA from resistant cells. Transfections will be performed into radiation sensitive human tumor cells and phenotypic selection will be performed to permit subsequent rescue of the DNA fragments. The identification of differentially expressed genes in radiation resistant cells will provide insight into the mechanisms of tumor radiation resistance and may be a future basis for tissue based predictive tests.

来自接受放射治疗的癌症患者的细胞系已经被 与临床结果和体外辐射反应相关。的 一些类似组织学的细胞，特别是来自 治疗未能局部控制他们的肿瘤，显示在体外 辐射生存参数的抗性类型，而其他人是一个 敏感型，表明辐射敏感性的遗传基础。 该项目的总体目标是识别可能与基因表达相关的基因。 与人类肿瘤的辐射抗性有关。使用 蛋白质分析技术（2-D凝胶电泳和 微测序）差异筛选/消减cDNA文库分析， 和载体介导的基因转移，一个多途径的战略计划， 在第一个目的，以确定和分离基因相关的 抗辐射表型。我们将检验这个假设， 细胞中的差异基因表达是辐射的原因， 敏感或耐辐射的肿瘤细胞表型。 在该项目的第二个目的中，两种识别基因的策略 与抵抗力相关的基因将被用来改变辐射反应， 敏感的肿瘤细胞的基因转移。第一 将使用附加型维持的载体策略， 第二种将使用粘粒策略， 利用来自抗性细胞的较大DNA片段进行基因转移。 将在对辐射敏感的人肿瘤细胞中进行转染 并进行表型选择以允许随后的拯救 DNA片段。差异表达基因的鉴定 抗辐射细胞将有助于深入了解肿瘤的机制 抗辐射性，并可能成为未来基于组织的预测的基础 试验.