Role of DDB2 in chemotherapeutic agents-induced apoptosis and platinum resistance

DDB2 在化疗药物诱导的细胞凋亡和铂类耐药中的作用

• 批准号: 8050717
• 负责人: Qien Wang
• 金额: $ 25.32万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8050717
• 关键词: Address; Apoptosis; Apoptotic; Binding; Biochemical; Biological Assay; Cancer cell line; Cell Line; Cell-Free System; Cells; Cisplatin; Clinical; Commit; DNA Damage; DNA-Binding Proteins; Development; Event; Exhibits; Flow Cytometry; Gene Proteins; Genetic Transcription; Human; Immunoprecipitation; Knowledge; Laboratories; Luciferases; Malignant Neoplasms; Malignant neoplasm of ovary; Measures; Mediating; Mismatch Repair; Molecular; Mus; Mutate; Names; Patients; Pharmaceutical Preparations; Platinum; Proteins; Reporting; Research; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Role; Sampling; Severe Combined Immunodeficiency; Signal Transduction; Solid Neoplasm; Suicide; Technology; Testing; Time; Tissues; Transfection; Translating; Tumor Tissue; Tumor-Derived; Western Blotting; Work; antitumor agent; base; cancer cell; cell injury; chemotherapeutic agent; chromatin immunoprecipitation; cytotoxicity; design; improved; in vivo; killings; knock-down; neoplastic; neoplastic cell; overexpression; pro-apoptotic protein; promoter; protein protein interaction; tumor

DESCRIPTION (provided by applicant): Cisplatin is one of the most potent anti-tumor agents, which displays clinical activity against a wide variety of solid tumors. The anti-neoplastic activity of cisplatin results from cisplatin- induced DNA damage, which further triggers cellular apoptosis. However, the effective use of cisplatin is limited by the development of cisplatin resistance in cancer cells. Damaged DNA binding protein (DDB2) has been reported to be able to bind UV- or cisplatin-induced DNA damage, and believed to have a correlation with the apoptotic event, especially the apoptosis induced by DNA damaging agents. However, the relationship between DDB2 and cisplatin- induced apoptosis needs to be validated and the mechanism needs to be explored. The specific hypothesis addressed in this proposal is that DDB2 mediates cisplatin-induced apoptosis through binding DNA damage to relay the apoptotic signals, and regulating pro- and anti-apoptotic proteins; as a result, DDB2 deficiency confers cancer cells resistance to cisplatin. The proposed work will utilize relevant biochemical, biophysical, immunological, cellular and molecular technologies mostly established and ongoing in the PI's laboratory to address the following specific aims: (1) To validate the role of DDB2 in cisplatin-induced apoptosis and the relationship between DDB2 and Bcl-2 in varying cancer cell lines and human tumor tissues; (2) To define the role of DDB2 recognition of cisplatin-induced DNA damage in triggering apoptosis; (3) To delineate the mechanism through which DDB2 down-regulates Bcl-2 level; (4) To reveal the relationship between DDB2 and p53 in the cisplatin-induced apoptosis; (5) To test the role of DDB2 in cisplatin sensitivity and cisplatin- induced apoptosis in vivo. The experimental focus of this proposal is on the relationship between DDB2 and apoptosis. Thus, the above-mentioned specific aims are designed to provide a comprehensive assessment of the regulatory function of DDB2 on cisplatin-induced apoptosis and the development of cisplatin resistance. PUBLIC HEALTH RELEVANCE: Cisplatin was first described as an anti-tumor agent in 1965 by Rosenberg. It has been in widespread clinical use for more than three decades to treat various malignant tumors. However, many human tumors have either intrinsic resistance to cisplatin or acquire resistance after the initial patient treatment. This has become a major problem limiting its effective use. Cisplatin kills tumor cells through interaction with cellular DNA and damaging it. The damaged DNA triggers programmed cell death pathway (apoptosis), and finally commits cancer cells to suicide. However, the cisplatin-resistant cancer cells hardly undergo apoptosis after cisplatin treatment due to multi-reasons. We have identified one protein, named damaged DNA binding protein 2 (DDB2), which can mediate cisplatin-induced apoptosis. In this proposal, we will further define how DDB2 facilitates cisplatin-induced programmed cell death in cisplatin-resistant cancer cells and how DDB2 sensitizes cisplatin-resistant tumors to cisplatin treatment. Ultimately, we hope to translate this knowledge into new strategies for increasing the cisplatin sensitivity and treating recalcitrant tumors.

描述（由申请人提供）：顺铂是最有效的抗肿瘤药物之一，它显示出针对各种实体瘤的临床活性。顺铂的抗塑性活性是由顺铂诱导的DNA损伤引起的，这进一步触发了细胞凋亡。然而，有效使用顺铂受到癌细胞中顺铂耐药性的发展的限制。据报道，受损的DNA结合蛋白（DDB2）能够结合UV-或顺铂诱导的DNA损伤，并被认为与凋亡事件有相关性，尤其是由DNA损害剂引起的凋亡。但是，需要验证DDB2与顺铂诱导的细胞凋亡之间的关系，需要探索该机制。该提案中解决的具体假设是，DDB2通过结合DNA损伤介导顺铂诱导的凋亡，从而传递凋亡信号，并调节促促凋亡蛋白。结果，DDB2缺乏赋予癌细胞对顺铂的抗性。拟议的工作将利用相关的生化，生物物理，免疫，细胞和分子技术，主要在PI实验室中建立和正在进行，以解决以下特定目的：（1）验证DDB2在顺倍肽诱导的凋亡以及DDB2之间的关系中的作用在不同的癌细胞系和人类肿瘤组织中的Bcl-2； （2）定义DDB2识别顺铂诱导的DNA损伤在触发凋亡中的作用； （3）描述DDB2下调Bcl-2水平的机制； （4）揭示了顺铂诱导的凋亡中DDB2和P53之间的关系； （5）测试DDB2在顺铂灵敏度和顺铂诱导的体内凋亡中的作用。该提议的实验重点是DDB2和凋亡之间的关系。因此，上述特定目的旨在对DDB2在顺铂诱导的细胞凋亡和顺铂耐药性的发展进行全面评估。 公共卫生相关性：顺铂于1965年首次被罗森伯格描述为抗肿瘤剂。三十年来，它一直在广泛的临床用途中治疗各种恶性肿瘤。但是，许多人类肿瘤在初次患者治疗后具有对顺铂的固有抵抗力或获得抗药性。这已成为限制其有效使用的主要问题。顺铂通过与细胞DNA相互作用并损坏肿瘤细胞。受损的DNA触发了编程的细胞死亡途径（凋亡），并最终将癌细胞自杀。然而，由于多季节，抗顺铂耐药细胞几乎不会在顺铂治疗后凋亡。我们已经确定了一种蛋白质，称为受损的DNA结合蛋白2（DDB2），该蛋白可以介导顺铂诱导的凋亡。在此提案中，我们将进一步定义DDB2如何促进顺铂诱导的抗铂耐药性癌细胞的程序性细胞死亡，以及DDB2如何使抗顺铂的耐药性肿瘤对顺铂治疗敏感。最终，我们希望将这些知识转化为增加顺铂敏感性和治疗顽固肿瘤的新策略。