MOLECULAR BIOLOGY OF B-CELL CHRONIC LYMPHOCYTIC LEUKEMIA

B 细胞慢性淋巴细胞白血病的分子生物学

• 批准号: 3198280
• 负责人: ELIZABETH W. NEWCOMB
• 金额: $ 15.22万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3198280
• 关键词: B lymphocyte; DNA; athymic mouse; chronic leukemia; gene expression; genetic manipulation; genetic mapping; human genetic material tag; human tissue; laboratory mouse; lymphatic tissue; lymphocytic leukemia; molecular cloning; northern blottings; nucleic acid sequence; oncogenes; southern blotting; transforming growth factors

Preliminary studies show transforming activity is associated with DNA from 6 cases of the advanced B cell malignancy, B cell chronic lymphocytic leukemia (B-CLL). The technique of DNA-mediated-gene-transfer was used to introduce high molecular weight DNA from B-CLL cells into mouse NIH 3T3 cells were inoculated into nude mice. Tumor formation is correlated with the transfer of DNA sequences which contain transforming activity. In this proposal we will clone the transforming gene(s) associated with advanced stages of B-CLL disease. The nude mouse tumorigenicity assay will monitor the cloning of the biologically active DNA fragment consisting of single copy human DNA will be used in Southern blot analysis to determine the pattern of gene arrangement in normal and malignant human lymphoid tissue. Northern blot analysis will determine expression of the gene(s) in mRNAs obtained from normal and malignant lymphoid tissues. To be cloned and its DNA sequence compared with the activated oncogene. The transforming DNA sequences will be analyzed by Southern blot using the active subclone to determine whether the genes are the same or different from one another. If the genes are the same, then fine restriction endonuclease mapping and DNA sequencing of the genes will determine the chromosomal change(s) involved in activation of the oncogene. The frequency of oncogene activation will be analyzed in B-CLL cases representative of different disease stages. This study will identify the transforming activity associated with B-CLL disease. Understanding the genetic mechanism of oncogene activation in B- CLL disease may have potential for improved patient diagnosis and treatment.

初步研究表明，转化活性与来自 6例晚期B细胞恶性肿瘤，B细胞慢性淋巴细胞 白血病（B-CLL）。 DNA介导的基因转移技术用于 将B-CLL细胞的高分子量DNA引入小鼠NIH 3T3中 将细胞接种到裸鼠中。 肿瘤形成与 含有转化活性的DNA序列的转移。 在这个 提案我们将克隆与高级相关的转化基因 B-CLL疾病的阶段。 裸小鼠肿瘤性测定将监测 由单个的生物活性DNA片段的克隆 复制人DNA将用于Southern印迹分析中以确定 正常和恶性的人淋巴组织中基因排列的模式。 北印迹分析将确定基因在mRNA中的表达 从正常和恶性淋巴组织获得。 克隆及其 与活化的癌基相比，DNA序列。 转化的DNA 将使用主动亚克隆来分析序列 确定基因是相同还是相同的。 如果 这些基因是相同的，然后精细的限制性核酸内切酶映射和DNA 基因的测序将确定涉及的染色体变化 在激活癌基因中。 致癌基因激活的频率将 在代表不同疾病阶段的B-CLL病例中进行分析。 这项研究将确定与B-CLL相关的转化活动 疾病。 了解B-中癌基活化的遗传机制 CLL疾病可能有可能改善患者诊断和 治疗。