DIFFERENTIALLY EXPRESSED GENE PRODUCTS IN GLIOMAS

胶质瘤中差异表达的基因产物

• 批准号: 2501908
• 负责人: Peter A Steck
• 金额: $ 10.17万
• 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 1997-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2501908
• 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; 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.

已经描述了人神经胶质瘤的肿瘤发生是通过 遗传损害的积累，最终导致激活 原始基因和/或肿瘤抑制基因功能的丧失。 该假设基于以前描述的研究 表皮生长因子受体基因的扩增和损失 与胶质体9、10和17相关的序列。 这 项目针对推定的本地化和识别 位于染色体上的胶质母细胞瘤相关肿瘤抑制（GTS）基因 10。几项调查表明染色体的缺失 10等位基因是在 胶质母细胞瘤（GBMS）。 为了进一步研究这一观察，我们有了 通过微细胞将人类10染色体插入两个胶质母细胞瘤细胞系 介导的染色体转移，并表现出巨大的损失 杂化细胞的致瘤表型。 杂交细胞表现出损失 它们在软琼脂糖中生长的能力，饱和密度降低 单层和未在裸体动物中形成肿瘤。 利用 这种功能方法证明了GTS基因在 染色体10。我们隔离GTS基因的长期目标涉及使用 在两种非纯正的排斥方法中：碎片染色体10，和 将各种定义的碎片重新插入到GBM单元中以找到区域 通过分析功能性肿瘤抑制，GTS基因。 我们的努力 特别将针对222年第10季度至10qter的区域 初步证据表明GTS基因可能存在。 此外， 将在cDNA表达文库上进行减法杂交 识别由于重复而差异表达的基因 将10染色体插入GBM细胞中。 差异表达 然后，基因产物的基因结构将被表征 表达模式，序列和基因结构，以最终 检查他们的功能作用。 此外，这两种方法可能是 合并以识别映射到特定染色体区域的基因 兴趣并差异表达。 这些结果应增加 我们对神经胶质涉及的特定遗传变化的理解 肿瘤发生，然后可能为我们提供新的诊断标记或特定的标记 人神经膜瘤治疗干预的靶标。