INHIBITION OF HUMAN ONCOGENE EXPRESSION BY INTERFERON

干扰素对人类癌基因表达的抑制

• 批准号: 3175180
• 负责人: ROBERT M FRIEDMAN
• 金额: $ 14.4万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1997-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3175180
• 关键词: DNA binding protein; aminoacid oxidase; athymic mouse; collagen; elastin; gene expression; genetic transcription; human genetic material tag; interferons; molecular oncology; neoplasm /cancer genetics; neoplastic transformation; oncogenes; tissue /cell culture

The enzyme lysyl oxidase functions as a suppressor of the phenotypic expression of the ras oncogene. NIH 3T3 mouse cells transformed by LTR-c-H-ras expressed very low levels of lysyl oxidase; expression was restored after interferon beta-mediated reversion. Revertants were retransformed by a lysyl oxidase antisense expression construct; the original transformed cells were reverted after transfection with a lysyl oxidase sense expression construct. Lysyl oxidase transcriptional regulatory sequences will be defined to investigate how the gene is switched off during ras transformation, and how reversion with interferon leads to the restoration of expression. Since demethylation of revertants caused retransformation and loss of lysyl oxidase expression, the control region may require methylation for activity. Positive and/or negative regulatory proteins that bind to the transcriptional regulatory control sequences, perhaps induced by either interferon or ras, will be identified and studied. The possible coordinate regulation of lysyl oxidase, collagen structural genes, and enzymes involved in collagen synthesis following ras transformation and reversion will be investigated. Lysyl oxidase expression in cell lines transformed by oncogenes other than ras will also be studied to reveal any similarities in method of action or interrelationships among ras and other oncogenes. If a relationship exists between lysyl oxidase activity and normal cell growth and behavior, understanding the regulatory mechanisms of this gene may provide opportunities for therapy: drugs that modify lysyl oxidase expression can be identified, and possibly in the future, gene replacement therapy could be considered. Cell lines of human cancers that arise from cells that produce collagen or elastin and lysyl oxidase will be examined for alterations in lysyl oxidase message expression and enzyme activity as well as collagen or elastin levels. If lysyl oxidase gene expression is decreased in any of these human cancers, expression experiments employing the human lysyl oxidase gene will be performed to attempt reversion. Lysyl oxidase has also been localized to some types of epithelia cells. Tumors arising from these tissues will be examined for any changes in lysyl oxidase expression. Normal tissue and cell lines derived from tumors of tissues that show a high incidence of mutation in the ras oncogene will also be examined to determine if there is a correlation between lysyl oxidase expression and transformation similar to that seen in the mouse fibroblast system. The relationship between immature collagen and elastin in cell matrix and possible changes in the binding/activity of growth-promoting or -controlling factors will also be studied.

赖氨酰氧化酶的功能是作为一种抑制剂， ras癌基因的表型表达。 NIH 3 T3小鼠细胞 LTR-c-H-ras转化表达非常低水平的赖氨酰 氧化酶;表达恢复后，干扰素β介导 逆转 回复突变体被赖氨酰氧化酶再转化 反义表达构建体;将原始转化细胞 用赖氨酰氧化酶正义表达转染后回复 构建体 赖氨酰氧化酶转录调控序列将 被定义为研究在ras过程中基因是如何被关闭的。 转化，以及干扰素逆转如何导致 恢复表达。 由于回复突变体的去甲基化 引起再转化和赖氨酰氧化酶表达的丧失， 控制区可能需要甲基化才能具有活性。 积极 和/或负调节蛋白，其结合到 转录调控序列，可能是由 干扰素或ras将被鉴定和研究。 的 赖氨酰氧化酶、胶原蛋白 结构基因和参与胶原蛋白合成的酶 随后将研究Ras转化和逆转。 赖氨酰氧化酶在癌基因转化细胞系中的表达 除了RAS之外，还将研究以揭示 行动方法或RAS与其他 致癌基因 如果赖氨酰氧化酶的活性 和正常的细胞生长和行为，了解调节 该基因的机制可能为治疗提供机会： 可以鉴定修饰赖氨酰氧化酶表达的药物， 在未来，基因替代疗法可能 考虑了 人类癌症的细胞系， 产生胶原蛋白或弹性蛋白和赖氨酰氧化酶将被检查 赖氨酰氧化酶信息表达和酶活性的改变 以及胶原蛋白或弹性蛋白水平。 如果赖氨酰氧化酶基因 在任何这些人类癌症中表达降低， 使用人赖氨酰氧化酶基因的实验将是 尝试恢复。 赖氨酰氧化酶也被 局限于某些类型的上皮细胞。 肿瘤来源于 将检查这些组织中赖氨酰氧化酶的任何变化 表情 来源于肿瘤的正常组织和细胞系 ras癌基因突变发生率高的组织 也将进行检查，以确定是否存在相关性 赖氨酰氧化酶的表达和类似于 在小鼠成纤维细胞系统中可见。 的关系 细胞基质中未成熟胶原和弹性蛋白及其可能的变化 在生长促进或控制因子的结合/活性中 也将被研究。