MOLECULAR ANALYSIS OF THE REGULATION PERTURBATION OF CELL CYCLE IN LUNG CANCER

肺癌细胞周期调控扰动的分子分析

• 批准号: 6504944
• 负责人: ROBERT A SCLAFANI
• 金额: $ 6.18万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6504944
• 关键词: athymic mouse; biomarker; carcinogenesis; cell growth regulation; clinical research; cyclin dependent kinase; cyclins; enzyme inhibitors; human tissue; immunocytochemistry; liposomes; lung neoplasms; molecular oncology; neoplasm /cancer genetics; neoplastic process; polymerase chain reaction; retinoblastoma protein; tissue /cell culture; transfection; western blottings

This proposal is part of a joint effort (SPORE) to provide better methods of clinical diagnosis, prognosis and treatments for lung cancer patients by studying the molecular basis of the disease. This project interacts with many of the other projects of this SPORE by sharing results and similar technologies to produce a synergistic effort. The project utilizes the human tissue procurement, tissue bank and animal cores. The focus of this project is on the role of cell cycle regulation in the etiology of lung cancer. The hypothesis is that precise, biochemical knowledge of the alterations of cell cycle regulatory molecules in cancer cells is essential for understanding the etiology of all cancers. This knowledge can then be exploited for solving the clinical problems of lung cancer patients, who have very little hope treatment. Molecular analysis focuses on the family of cyclin-dependent kinase (CDKs) and inhibitors (CDIs) and important CDK substrates such as the Rb (Retinoblastoma) tumor suppressor. A biochemical analysis of cyclin D1- CDK6 complexes and two CDIs, the MTS1 and mTS2 (multiple tumor suppressor) genes, is being performed in order to investigate the mechanism of perturbed cell cycle regulation in NSCLC cells. This molecular analysis can also reveal additional molecules altered in NSCLC and provide the basis for potential prognosis and therapy. Potential gene therapies and a test of the hypothesis that overexpressed cyclin D1 and loss of MTS1-2 is important for aberrant proliferation in NSCLC is accomplished by the use of recombinant DNA technologies, which employ antisense cyclin D1 retroviruses, plasmids and synthetic oligodeoxynucleotides, as well as MTS1-2 expression plasmids. Liposome technologies are used to deliver the plasmids and synthetic oligodeoxynucleotides to NSCLC cells in vitro and in vivo. Nude rodents with xenografts of human tumor cells are used to test the efficacy of these genetic manipulations and therapies in vivo. Molecular analysis of human tumors, dysplastic tissues and sputum samples are screened with a large panel of molecular markers, including but is not limited, to, cyclin D1, CDKs, CDIs and Rb using PCR, PCR-SSCP, immunoblots and immunohistochemical analyses. In this manner, a systematic study of any genetic changes that occur during tumor progression is done. These genetic changes can then be correlated with pathological changes in the cells to produce a molecular model for tumor progression. Dysplastic human cells that have cell cycle alterations are transformed with a variety of oncogenes as an assay of their state of progression to provide direct experimental evidence for such a molecular model. This molecular model has the potential to be used as a monitor of tumor progression in patients by providing oncologists with both prognostic and diagnostic information.

该建议是提供更好方法的联合努力（孢子）的一部分 通过临床诊断，预后和肺癌患者的治疗 研究该疾病的分子基础。 这个项目与 该孢子的其他许多项目通过共享结果和类似 产生协同努力的技术。 该项目利用 人体组织采购，组织库和动物核心。 该项目的重点是细胞周期调节在 肺癌病因。 假设是精确的生化 了解癌症细胞周期调节分子的改变 细胞对于理解所有癌症的病因至关重要。 这 然后可以利用知识来解决肺的临床问题 癌症患者，希望治疗很少。 分子分析侧重于细胞周期蛋白依赖性激酶（CDKS）家族 和抑制剂（CDI）和重要的CDK底物（例如RB） （视网膜母细胞瘤）肿瘤抑制剂。 细胞周期蛋白D1-的生化分析 CDK6复合物和两个CDI，MTS1和MTS2（多个肿瘤抑制剂） 正在执行基因，以研究 NSCLC细胞中的扰动细胞周期调节。 该分子分析 还可以揭示NSCLC中改变的其他分子并提供基础 潜在的预后和治疗。 潜在的基因疗法和对过表达的假设的检验 Cyclin D1和MTS1-2的丧失对于异常增殖很重要 NSCLC是通过使用重组DNA技术来完成的， 使用反义细胞周期蛋白D1逆转录病毒，质粒和合成 寡脱氧核苷酸以及MTS1-2表达质粒。 脂质体 技术用于传递质粒和合成 在体外和体内对NSCLC细胞的寡脱氧核苷酸。 裸啮齿动物 使用人类肿瘤细胞的异种移植物用于测试这些细胞的功效 体内的遗传操作和疗法。 人肿瘤，发育不良组织和痰样品的分子分析 用大型分子标记筛选，包括但不是 使用PCR，PCR-SSCP，免疫印迹限制，至Cyclin D1，CDK，CDIS和RB 和免疫组织化学分析。 以这种方式，对 在肿瘤进展过程中发生的任何遗传变化都是进行的。 这些 然后，遗传变化可以与病理变化相关 细胞产生用于肿瘤进展的分子模型。 发育不良的人 具有细胞周期改变的细胞通过多种 癌基因作为其进展状态的测定，以提供直接 这种分子模型的实验证据。 这个分子模型具有 用作患者肿瘤进展监测的潜力 为肿瘤学家提供预后和诊断信息。