HYPOXIA STRESS GENES IN VITRO AND IN SOLID TUMORS

体外和实体瘤中的缺氧应激基因

• 批准号: 6300485
• 负责人: ROBERT M SUTHERLAND
• 金额: $ 21.86万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-14 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6300485
• 关键词: antitumor antibody; cis platinum compound; drug resistance; gene expression; head /neck neoplasm; heme oxygenase; hypoxia; immunocytochemistry; in situ hybridization; neoplasm /cancer genetics; neoplastic cell; polymerase chain reaction; stress proteins; tissue /cell culture

Deficiencies in tumor blood supply establish gradients of oxygen, glucose, growth regulators, and other substances along diffusion paths radiating from blood vessels. These stress environments represent physiological differences between tumor and normal tissue. SRI researchers have shown that hypoxic stress in human and rodent tumor cells induces the synthesis of a set of proteins called oxygen regulated proteins or ORPs. It is hypothesized that genes regulated by hypoxic stress are important participants in malignant progression. To investigate this hypothesis, SRI proposes to measure the expression of specific ORPs in tumor tissue and to develop a model to test the relationship of expression of an ORP with resistance to therapy. This information will permit the creation of novel strategies for determining the prognosis and treatment of some human cancers. Two approaches will be used to identify hypoxic regions of tumor tissue (FaDu, SQ-20B, and A431 human squamous carcinoma cells) and establish correlations with ORP expression: (1) antibodies to a fluorinated bioreductive drug will be used as a positive control to visualize hypoxic cells in tumor models in vitro, and the expression of selected ORPs (e.g., heme oxygenase-1 [HO-1; ORP 33], metallothionein IIA [MT-IIA; ORP 7], and vascular endothelial growth factor [VEGF]) will be detected by in situ hybridization and immunohistochemistry; and (2) similar experiments will be performed with human tumor xenografts. SRI's preliminary studies indicate that human MT-IIA is a suitable ORP for investigating the development of therapeutic resistance caused by hypoxia and reoxygenation. We will use transient and stable transfectants of human squamous carcinoma cells to create multicellular tumor spheroid models for investigating (1) a hypoxia-responsive MT-IIA 5'-regulatory region and (2) the effect of MT-IIA expression on resistance to cisplatin. These experiments are directed toward creating an in vitro tumor model of the human MT-IIA (ORP 7) 5'-regulatory region and coding sequence for investigation of the effects of hypoxic and glucose stress on a clinically relevant hypoxic stress protein. This model will provide a test system for inducible drug resistance associated with hypoxia and reoxygenation.

肿瘤血液供应的不足建立了氧气、葡萄糖、 生长调节剂和其他沿扩散路径辐射的物质 来自血管。这些压力环境代表了生理上的 肿瘤组织与正常组织之间的差异。SRI研究人员表明 人和啮齿动物肿瘤细胞的低氧应激诱导合成 一组被称为氧调节蛋白或ORPs的蛋白质。它是 假设受低氧应激调控的基因很重要 恶性进展的参与者。为了研究这一假设， SRI建议测量肿瘤组织中特定ORPs的表达 并开发了一个模型来检验ORP表达之间的关系 对治疗产生抗拒。该信息将允许创建 确定某些人类预后和治疗的新策略 癌症。 将使用两种方法来识别肿瘤组织的缺氧区 (FaDu、SQ-20B和A431人鳞癌细胞)并建立 与ORP表达的相关性：(1)氟化的抗体 生物还原药物将作为阳性对照用于显示低氧 体外肿瘤模型中的细胞，以及选定的ORPs的表达(例如， 血红素加氧酶-1[HO-1；ORP33]，金属硫蛋白IIA[MT-IIA；ORP7]，以及 血管内皮细胞生长因子[VEGF])将通过原位检测 杂交和免疫组织化学；以及(2)类似的实验将 用人肿瘤异种移植瘤进行手术。 SRI的初步研究表明，人MT-IIA是一种适合于 低氧引起的治疗抵抗的研究进展 和复氧作用。我们将使用瞬时和稳定的转染体 人鳞癌细胞形成多细胞肿瘤球体 研究(1)低氧反应MT-IIA5‘-调节的模型 (2)MT-IIA表达对顺铂耐药的影响。 这些实验旨在建立一种体外肿瘤模型 人MT-IIA(ORP7)5‘-调控区及其编码序列 低氧和葡萄糖应激对临床的影响 相关的低氧应激蛋白。该模型将提供一种测试系统 用于与缺氧和复氧相关的诱导性耐药性。