Mitochondrial DNA in Mutations in Prostate Cancer

前列腺癌突变中的线粒体 DNA

• 批准号: 6782685
• 负责人: John A. Petros
• 金额: $ 24.72万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6782685
• 关键词: apoptosis; athymic mouse; carcinogenesis; cell cycle; cell line; cell proliferation; clinical research; cytoplasm; flow cytometry; free radical oxygen; gene mutation; human tissue; hybrid cells; mitochondrial DNA; neoplastic growth; nucleic acid sequence; phenotype; prostate neoplasms; transfection; tumor promoters

DESCRIPTION (provided by applicant): Mitochondrial DNA (mtDNA) mutations occur with high frequency in a variety of human tumors but the functional consequences of such mutations are unknown. The overall goal of our proposal is to understand how mtDNA mutations affect the malignant potential of cancer cells. We hypothesize that mtDNA mutations enhance the survival and proliferation while decreasing apoptosis of those tumors cells that happen to acquire them. We further propose that mtDNA mutations increase the mitochondrial production of reactive oxygen species (ROS) and that this increased ROS production is mitogenic, enhancing proliferation and decreasing apoptosis. Preliminary data from our lab shows that one such mtDNA mutation is indeed capable of enhancing the growth of human prostate cancer xenografts. In order to test this hypothesis 3 specific aims are proposed. First we will generate the laboratory constructs required to test the hypothesis. Specifically, we will create cytoplasmic hybrids (cybrids) that combine the nucleus of well-characterized prostate cancer cell lines with cytoplasm (and therefore mitochondria) containing either mutant or wild type mtDNA. Second, we will determine whether specific mutations affect the malignant phenotype, measuring cellular proliferation, apoptosis, cell cycle progression and tumorigenesis. Finally, we will determine the role of ROS in the observed phenotypic alterations, establishing whether ROS are a functional intermediate between mtDNA mutation and enhanced tumor growth. The accomplishment of these aims will allow us to determine whether mtDNA mutations enhance the malignant phenotype and whether such an alteration is due to increased ROS. In addition, we will generate cells, tumors and animal models that will allow the future study of the role of mtDNA in cancer by other investigators and aid in the pre-clinical screening of potential anti-neoplastics directed at tumors that harbor mtDNA mutations.

描述（由申请人提供）：线粒体DNA （mtDNA）突变在各种人类肿瘤中发生频率很高，但此类突变的功能后果尚不清楚。我们建议的总体目标是了解mtDNA突变如何影响癌细胞的恶性潜能。我们假设mtDNA突变增强了肿瘤细胞的存活和增殖，同时减少了碰巧获得它们的肿瘤细胞的凋亡。我们进一步提出，mtDNA突变增加了线粒体活性氧（ROS）的产生，这种增加的ROS产生是有丝分裂的，促进增殖和减少凋亡。我们实验室的初步数据表明，一种这样的mtDNA突变确实能够促进人类前列腺癌异种移植物的生长。为了验证这一假设，提出了3个具体目标。首先，我们将生成检验假设所需的实验室结构。具体来说，我们将创造细胞质杂交（cybrids），将具有良好特征的前列腺癌细胞系的细胞核与含有突变型或野生型mtDNA的细胞质（因此是线粒体）结合起来。其次，我们将确定特定突变是否影响恶性表型，测量细胞增殖、凋亡、细胞周期进展和肿瘤发生。最后，我们将确定ROS在观察到的表型改变中的作用，确定ROS是否是mtDNA突变和肿瘤生长增强之间的功能性中介。这些目标的实现将使我们能够确定mtDNA突变是否会增强恶性表型，以及这种改变是否由于ROS增加。此外，我们将生成细胞、肿瘤和动物模型，这将允许其他研究人员在未来研究mtDNA在癌症中的作用，并有助于针对含有mtDNA突变的肿瘤的潜在抗肿瘤药物的临床前筛选。