ISOLATION OF A SELECTED DRUG RESISTANT GENE

选定的耐药基因的分离

• 批准号: 2683675
• 负责人: Lori A Hazlehurst
• 金额: $ 3.05万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-18 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2683675
• 关键词: MCF7 cell; adenosine triphosphate; analog; antineoplastics; artificial chromosomes; chromosome walking; complementary DNA; drug metabolism; drug resistance; fluorescent in situ hybridization; gene expression; genetic library; genetic mapping; high performance liquid chromatography; mass spectrometry; membrane transport proteins; mitoxantrone; neoplasm /cancer pharmacology; nuclear magnetic resonance spectroscopy; polymerase chain reaction; protein purification; protein sequence

The emergence of tumors refractory to further chemotherapy is a major obstacle for the successful treatment of cancer. Overexpression of MDR1 has been associated with clinical multidrug resistance. However, clinical trials have indicated that other as yet unidentified mechanisms occur in clinical multidrug resistance. This laboratory has selected mitoxantrone resistant human cell lines which is associated with an energy dependent drug efflux but is not related to the overexpression of known drug transporters, MDR1 and MRP. These cell lines provide a model of a novel multidrug resistant phenotype. We have recently determined by chromosome transfer that the resistance associated with mitoxantrone selection is due to a dominant genetic event. This provides the rationale for the cloning of the gene responsible for mitoxantrone selected resistance and is the goal of this proposal. A major undertaking of this proposal is the development of probes to screen a cDNA library. We have invoked strategies that utilize photoaffinity analogs of mitoxantrone as well as ATP photoaffinity analogs to identify putative proteins involved in resistance. Purification of proteins which are tagged in resistant cell lines by the photo affinity analogs will ultimately allow us to develop antibodies for immunoscreening a cDNA expression library. Purified proteins will also be sequenced which will allow us to design degenerate oligos for screening of a cDNA library. Finally, we propose that if we are unable to identify probes as outlined in this proposal we will proceed to physically map the gene. This will be accomplished by utilizing YAC libraries generated from chromosome transfer hybrids to clone the mitoxantrone selected resistant gene.

肿瘤对进一步化疗的难治性的出现是一个主要问题

项目成果

Multiple mechanisms confer drug resistance to mitoxantrone in the human 8226 myeloma cell line.

• 发表时间: 1999-03
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: L. Hazlehurst;Nils E. Foley;M. Gleason-Guzman;M. Hacker;A. Cress;L. Greenberger;M. C. D. Jong;W. Dalton