DIFFERENTIALLY EXPRESSED GENE PRODUCTS IN HUMAN GLIOMAS

人类胶质瘤中差异表达的基因产物

• 批准号: 3200520
• 负责人: Peter A Steck
• 金额: $ 18.09万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-04-01 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3200520
• 关键词: athymic mouse; chromosome deletion; gene expression; genetic library; genetic manipulation; genetic mapping; glioblastoma multiforme; glioma; human tissue; hybrid cells; in situ hybridization; neoplasm /cancer genetics; neoplasm /cancer remission /regression; northern blottings; nucleic acid sequence; nucleic acid structure; polymerase chain reaction; radiotracer; tumor suppressor genes

The oncogenesis of human gliomas has been described to occur through the accumulation of genetic damage that eventually leads to the activation of proto-oncogenes and/or the loss of function of tumor suppressor genes. This hypothesis is based on previous studies that have described amplification of epidermal growth factor receptor gene and losses of sequences associated with chromosomes 9, 10, and 17 in gliomas. This project is directed at the localization and identification of a putative glioblastoma-associated tumor suppressor (GTS) gene located on chromosome 10. Several investigations have demonstrated that deletion of chromosome 10 alleles are the most frequent genetic alteration observed in glioblastomas (GBMs). To further examine this observation, we have re- inserted human chromosome 10 into two glioblastoma cell lines by micro-cell mediated chromosomal transfer and have demonstrated a dramatic loss of the tumorigenic phenotype of the hybrid cells. The hybrid cells exhibit a loss of their ability to grow in soft agarose, a decreased saturation density in monolayer, and failure to form tumor in nude animals. The utilization of this functional method demonstrates the presence of a GTS gene on chromosome 10. Our long-term goal to isolate the GTS gene involves the use of two non-mutually exclusive approaches: to fragment chromosome 10, and to re-insert the various defined pieces into GBM cells to regionally locate the GTS gene by assaying for functional tumor suppression. Our efforts will particularly be directed towards the region 10q22 to 10qter, where preliminary evidence suggests the GTS gene may reside. Additionally, subtractive hybridizations will be performed on cDNA expression libraries to identify genes which are differentially expressed as a result of re- insertion of chromosome 10 into GBM cells. The differentially expressed gene products will then be characterized for their gene structure, expression patterns, sequence, and gene structure, in order to eventually examine their functional roles. Furthermore, these two approaches can be combined to identify genes which map to specific chromosomal regions of interest and are differentially expressed. These results should increase our understanding of the specific genetic alterations involved in glial oncogenesis and may then provide us with new diagnostic markers or specific targets for therapeutic intervention in human gliomas.

据描述，人类神经胶质瘤的肿瘤发生是通过 基因损伤的积累最终导致激活 原癌基因和/或抑癌基因功能丧失。 这个假设是基于之前的研究，这些研究描述了 表皮生长因子受体基因扩增和缺失 与神经胶质瘤中的 9、10 和 17 号染色体相关的序列。 这 项目旨在定位和识别假定的 位于染色体上的胶质母细胞瘤相关肿瘤抑制（GTS）基因 10. 多项研究表明，染色体缺失 10 个等位基因是观察到的最常见的遗传改变 胶质母细胞瘤（GBM）。 为了进一步检验这一观察结果，我们重新 通过微细胞将人类10号染色体插入两个胶质母细胞瘤细胞系中 介导的染色体转移并已证明 杂交细胞的致瘤表型。 混合电池表现出损失 它们在软琼脂糖中生长的能力，饱和密度降低 单层，在裸动物中不能形成肿瘤。 利用 该功能方法证明了 GTS 基因的存在 10 号染色体。我们分离 GTS 基因的长期目标涉及使用 两种非互斥的方法：将 10 号染色体片段化，以及 将各种定义的片段重新插入 GBM 细胞中以进行区域定位 通过检测功能性肿瘤抑制来检测 GTS 基因。 我们的努力 将特别针对 10q22 至 10qter 区域，其中 初步证据表明 GTS 基因可能存在。 此外， 将在 cDNA 表达文库上进行消减杂交 识别因重新调整而差异表达的基因 将 10 号染色体插入 GBM 细胞中。 差异表达的 然后将表征基因产物的基因结构， 表达模式、序列和基因结构，以便最终 检查他们的功能角色。 此外，这两种方法可以 结合起来识别映射到特定染色体区域的基因 兴趣并有不同的表达。 这些结果应该会增加 我们对神经胶质细胞特定遗传改变的理解 肿瘤发生，然后可能为我们提供新的诊断标记物或特定的 人类神经胶质瘤治疗干预的目标。