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 晚期B细胞恶性肿瘤6例，B细胞慢性淋巴细胞性白血病6例，恶性淋巴细胞性白血病6例。 白血病（B-CLL）。 利用DNA介导的基因转移技术， 将B-CLL细胞高分子量DNA导入小鼠NIH 3T3 将细胞接种到裸鼠中。 肿瘤的形成与 含有转化活性的DNA序列的转移。 在这 建议我们将克隆与晚期乳腺癌相关的转化基因， B-CLL疾病的阶段。 裸鼠致瘤性试验将监测 克隆由单个DNA片段组成的生物活性DNA片段， DNA拷贝将用于Southern印迹分析，以确定 人类正常和恶性淋巴组织的基因排列模式。 北方印迹分析将确定mRNA中基因的表达 从正常和恶性淋巴组织中获得。 被克隆， DNA序列与激活的癌基因的比较。 转化dna 使用活性亚克隆通过Southern印迹分析序列， 确定这些基因是相同还是不同。 如果 基因是相同的，然后精细的限制性内切酶图谱和DNA 基因测序将确定所涉及的染色体变化 致癌基因的激活。 癌基因激活的频率 在代表不同疾病阶段的B-CLL病例中进行分析。 本研究将确定与B-CLL相关的转化活性 疾病 了解B-细胞癌基因激活的遗传机制 CLL疾病可能具有改善患者诊断的潜力， 治疗