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: 6103065
• 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: 1999
• 资助国家: 美国
• 起止时间: 1999-05-13 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6103065
• 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.

肿瘤血液供应中的缺陷建立了氧，葡萄糖， 生长调节剂和其他物质沿着扩散路径辐射 从血管中。 这些压力环境代表了生理上的 肿瘤与正常组织的区别 SRI研究人员表示， 人类和啮齿类动物肿瘤细胞中的缺氧应激诱导了 一组叫做氧调节蛋白或ORP的蛋白质。 是 假设受缺氧应激调节的基因很重要 参与恶性进展。 为了研究这个假设， SRI建议测量肿瘤组织中特定ORP的表达 并建立了一个模型来测试一个ORP的表达关系， 对治疗有抵抗力 这些信息将允许创建 用于确定某些人类肿瘤的预后和治疗的新策略 癌的 两种方法将用于确定肿瘤组织的缺氧区域 (FaDu SQ-20 B和A431人鳞状细胞癌细胞），并建立 与ORP表达的相关性：（1）氟化抗体 生物还原性药物将用作阳性对照，以观察缺氧 体外肿瘤模型中的细胞，以及所选ORP（例如， 血红素加氧酶-1 [HO-1; ORP 33]，金属硫蛋白IIA [MT-IIA; ORP 7]，和 血管内皮生长因子[VEGF]）将通过原位 杂交和免疫组织化学;和（2）类似的实验将 用人肿瘤异种移植物进行。 SRI的初步研究表明，人MT-IIA是一种合适的ORP， 研究缺氧引起的治疗抗性的发展 和复氧。 我们将使用瞬时和稳定的转染子， 人鳞状细胞癌细胞产生多细胞肿瘤球体 用于研究（1）低氧响应性MT-IIA 5 '-调节的模型 （2）MT-IIA表达对顺铂耐药的影响。 这些实验旨在建立体外肿瘤模型， 人MT-IIA（ORP 7）5 ′-调控区和编码序列 低氧和葡萄糖应激对急性心肌梗死临床疗效的影响 相关低氧应激蛋白。 该模型将提供一个测试系统 与缺氧和复氧相关的诱导性耐药性。