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

该提案是联合努力（SPORE）的一部分，旨在提供更好的方法 对肺癌患者的临床诊断、预后和治疗有重要意义。 研究这种疾病的分子基础 该项目与 许多其他项目的这个孢子通过分享成果和类似的 技术，以产生协同效应。 该项目利用 人体组织获取、组织库和动物核心。 该项目的重点是细胞周期调控在细胞周期调控中的作用。 肺癌的病因。 假设是精确的，生化的 了解癌症中细胞周期调节分子的改变 细胞对于了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部的临床问题， 癌症患者，他们几乎没有治疗的希望。 分子分析主要集中在细胞周期蛋白依赖性激酶（CDKs）家族 和抑制剂（CDIs）和重要的CDK底物，如Rb （视网膜母细胞瘤）肿瘤抑制因子。 细胞周期蛋白D1的生化分析 CDK 6复合物和两种CDIs，MTS 1和mTS 2（多肿瘤抑制因子） 基因，正在进行，以调查的机制， 在NSCLC细胞中干扰细胞周期调节。 这种分子分析 还可以揭示NSCLC中改变的其他分子， 用于潜在的预后和治疗。 潜在的基因疗法和一个假设的测试，过度表达 细胞周期蛋白D1和MTS 1 -2的缺失对于细胞异常增殖很重要 NSCLC是通过使用重组DNA技术实现的， 使用反义细胞周期蛋白D1逆转录病毒、质粒和合成的 寡脱氧核苷酸以及MTS 1 -2表达质粒。 脂质体 技术被用于递送质粒和合成的 在体外和体内，寡脱氧核苷酸对NSCLC细胞的作用。 裸啮齿动物 与人肿瘤细胞的异种移植物一起用于测试这些 基因操作和体内治疗。 人类肿瘤、异型增生组织和痰样本的分子分析 用大量的分子标记进行筛选，包括但不限于 应用PCR、PCR-SSCP、免疫印迹等方法， 和免疫组织化学分析。 通过这种方式，系统地研究 在肿瘤进展过程中发生的任何遗传变化都是如此。 这些 然后，遗传变化可以与组织中的病理变化相关。 细胞以产生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被各种 癌基因作为其进展状态的测定，以提供直接的 实验证据证明了这种分子模型。 这个分子模型 作为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。

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.