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

这项提议是提供更好方法的联合努力的一部分。 对肺癌患者的临床诊断、预后和治疗的影响 研究这种疾病的分子基础。该项目与 这个孢子的许多其他项目通过分享结果和类似的 产生协同效应的技术。该项目利用了 人体组织采购、组织库和动物核心。 这个项目的重点是细胞周期调节在细胞周期中的作用。 肺癌的病因学。假设是精确的，生化的 细胞周期调控分子在癌症中变化的认识 细胞对于理解所有癌症的病因是必不可少的。这 这样就可以利用知识来解决肺部的临床问题。 癌症患者，他们的治疗希望非常渺茫。 分子分析的重点是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，如Rb (视网膜母细胞瘤)肿瘤抑制因子。细胞周期蛋白D_1的生化分析 CDK6复合体和两个CDI，MTS1和MTS2(多肿瘤抑制因子) 基因，正在进行研究，以研究 干扰了非小细胞肺癌细胞的细胞周期调节。这项分子分析 也可以揭示非小细胞肺癌中改变的额外分子，并提供基础 用于潜在的预后和治疗。 潜在的基因疗法和对过度表达假说的检验 细胞周期蛋白D1和MTS1-2的缺失在卵巢癌细胞异常增殖中起重要作用 NSCLC是通过使用重组DNA技术来完成的，该技术 利用反义细胞周期蛋白D1逆转录病毒、质粒和人工合成 寡脱氧核苷酸，以及MTS1-2表达载体。脂质体 技术被用来传递质粒和合成 寡脱氧核苷酸对非小细胞肺癌细胞的体内外杀伤作用。裸鼠 用人体肿瘤细胞的异种移植来测试这些药物的疗效 体内的基因操作和治疗。 人体肿瘤、异型增生组织和痰标本的分子分析 用一大批分子标记进行筛选，包括但不是 利用PCR、PCR-SSCP、免疫印迹技术对细胞周期蛋白D1、CDKs、CDI和Rb进行限制 和免疫组织化学分析。通过这种方式，系统地研究了 在肿瘤进展过程中发生的任何基因变化都是完成的。这些 然后，基因变化可以与心脏疾病的病理变化相关联 细胞产生一个肿瘤进展的分子模型。发育不良的人类 有细胞周期改变的细胞用各种不同的 癌基因作为对其进展状态的分析提供了直接的 这种分子模型的实验证据。这个分子模型有 通过以下方法监测患者肿瘤进展的可能性 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.