SPECIFIC INHIBITION OF HER2/NEU TRANSCRIPTION ELONGATION

描述：HER2/neu癌基因似乎在 多种人类癌症的发生和发展，包括 大约25%的非小细胞肺癌(NSCLC)，主要是 美国男性和女性的死亡原因。这一总体目标 项目是寻找新的方法通过以下方式特异性地抑制HER2/neu的表达 开发寡核苷酸(ODN)-百菌清(CHL)结合物 通过三链DNA与HER2/neu基因定点结合 并通过CHL在特定的鸟嘌呤碱基上导致DNA烷基化。 具体地说，本申请中概述的工作将完成 以下目标：1)表征百菌清的偶联能力 β(天然)和α(核酸酶抗性修饰)异构体ODN HER-2/neu基因DERET位点特异性DNA烷基化；2)特征 与百菌清结合的寡核苷酸与HER/neu基因结合的能力 抑制HER-2/neu基因转录延长的体外实验和cDNA实验 表达载体导入HeLa细胞和NIH3T3细胞；3) 显示染色质中的ODN结合和位点特异性DNA修饰 表达HER-2/neu基因的活的NSCLC细胞；4)表征 腺病毒介导ODN摄取和细胞核定位的能力 当腺病毒-ODN复合体与化学连接物形成时，NSCLC细胞。 5)确定以最佳方式递送的ODN-CHL结合物的能力 抑制HER-2/neu基因表达逆转人卵巢癌恶性表型 人非小细胞肺癌组织培养及啮齿动物模型的建立。的具体目标 此应用程序旨在解决以下主要障碍 成功开发了一种基于ODN的位点特异性DNA结合药物。这个 成功完成这些具体目标将带来无可估量的 对基因特异性DNA结合化合物设计的见解。这个 HER-2/neu基因特异性抗基因化合物的开发将提供一种 大量关于HER-2/neu基因在HER-2/neu基因中的作用 非小细胞肺癌的发生和发展并可能导致新的治疗方法 HER-2/neu基因表达非小细胞肺癌等肿瘤的途径

DESCRIPTION: The HER2/neu oncogene appears to play an important role in the initiation and progression of many types of human cancer, including approximately 25 percent of non-small cell lung cancer (NSCLC), the leading cause of death in both men and women in the U.S. The overall goal of this project is to find novel ways to specifically inhibit HER2/neu expression by developing oligonucleotide (ODN) - chlorambucil (CHL) conjugates that will bind in a site-specific manner to the HER2/neu gene by triplex DNA formation and lead to DNA alkylation at specific guanine bases by the CHL. Specifically, the work outlined in this application will accomplish the following goals: 1) Characterize the ability of chlorambucil-conjugated beta (natural) and alpha (nuclease resistant modification) anomeric ODNs to diret site-specific DNA alkylation in the HER-2/neu gene; 2) Characterize the ability of chlorambucil-conjugated ODNs to bind to the HER/neu gene and inhibit HER-2/neu gene transcription elongation in vitro and in a cDNA expression plasmid transfected into HeLa cells and NIH3T3 cells; 3) Demonstrate ODN binding and site-specific DNA modification in the chromatin of living NSCLC cells that express the HER-2/neu gene; 4) Characterize the ability of adenoviruses to mediate ODN uptake and nuclear localization in NSCLC cells when adenovirus-ODN complexes are formed with a chemical linker. 5) Determine the ability of optimally delivered ODN-CHL conjugates to inhibit HER-2/neu gene expression and reverse the malignant phenotype in tissue culture and rodent models of human NSCLC. The specific objectives of this application are designed to address the major obstacles to the successful development of an ODN-based site-specific DNA binding drug. The successful completion of these Specific Aims will lead to invaluable insights into the design of gene-specific DNA binding compounds. The development of a HER-2/neu gene specific anti-gene compound will provide a great deal of information about the role of the HER-2/neu gene in the initiation and progression of NSCLC and may lead to novel treatment approaches for HER-2/neu gene expressing cancers such as NSCLC.