CHROMOSOME ALTERATIONS AND PROTO-ONCOGENES TRANSPOSITION IN CARCINOGENESIS

致癌过程中的染色体改变和原癌基因转座

• 批准号: 3853472
• 负责人: N C POPESCU
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3853472
• 关键词: Burkitt's lymphoma; Epstein Barr virus; Papillomavirus; Tay Sachs disease; cell line; cervix neoplasms; chemical carcinogenesis; chromosome disorders; chromosome translocation; fluorescence; gel electrophoresis; gene expression; genetic mapping; genetic recombination; human tissue; in situ hybridization; linkage mapping; neoplasm /cancer genetics; nucleic acid probes; nucleic acid sequence; oncogenes; protooncogene; thyroid hormone binding protein; transfection; viral carcinogenesis; virus genetics

Non-isotopic in situ hybridization is a powerful molecular approach for detecting and localizing specific nucleic acid sequences within interphase nuclei or chromosomes. Single viral copies were detected by fluorescent signals after hybridization with biotinylated virus DNA probes. This has a practical significance for detecting DNA and RNA viruses in malignant and premalignant lesions. On a cervical carcinoma cell line (C4-1), human papillomavirus-18 (HPV-18) sequences were localized at region 8q2l on an 8;12 rearranged chromosome. In a Burkitt's lymphoma cell line, Epstein-Barr virus sequences were mapped on chromosome 2pl3 adjacent to a viral modification site. In both lines viral integration corresponded with the location of a fragile site. A nontumorigenic line (CX16-2) derived from exocervical epithelial cells transfected with recombinant HPV-16, hoarbor viral sequences on chromosome 2 near ets-2 gene. The ets-2 specific m-RNA level was elevated in the absence of structural gene alterations. However, no linkage between ets-2 and HPV-16 sequences was established by pulse field gel electrophoresis using several rarecutting restriction enzymes. Thus, HPV sequences can influence, from a distance, proto-oncogene expression. Several HPV-16 integration sites exhibited an aberrant late replication pattern. Incomplete chromatin condensation and recombination are consequences of the replication junction that flank late replicating DNA and can explain the origin of chromosomal changes associated with the cell's immortality. Neu proto-oncogene, the rat homolog of the human erb-B-2-gene, was neither amplified nor overexpressed in two rat mammary cell lines neoplastically transformed in vitro by a chemical carcinogen. A cDNA for the gene that encodes a human cytosolic thyroid hormone binding protein (p58) was mapped by in situ hybridization to 15q24-25. This localization may serve as a useful marker for Tay-Sachs disease and will permit assessment of the effect chromosome alterations involving this region have on human malignancies.

非同位素原位杂交是一种强有力的分子方法， 检测和定位特定核酸序列， 间期核或染色体。 检测到单病毒拷贝， 与生物素化病毒DNA杂交后的荧光信号 probes. 这对DNA和RNA的检测具有实际意义 病毒在恶性和癌前病变。 子宫颈癌 细胞系（C4-1），人乳头瘤病毒-18（HPV-18）序列， 位于8;12重排染色体上的8 q2 l区域。 中 伯基特淋巴瘤细胞系，爱泼斯坦-巴尔病毒序列被定位在 与病毒修饰位点相邻的染色体2 p13。 在两个品系中 病毒整合对应于脆性位点的位置。 一 来源于外表皮细胞的非致瘤细胞系（CX 16 -2） 用重组HPV-16转染， 2号染色体靠近ets-2基因。 ets-2特异性mRNA水平为 在没有结构基因改变的情况下升高。 但没有 用脉冲电场建立了ets-2和HPV-16之间的连锁关系 使用几种罕见的限制酶进行凝胶电泳。 因此，在本发明中， HPV序列可以从远处影响原癌基因的表达。 几个HPV-16整合位点表现出异常的晚期复制 格局 不完全染色质凝聚和重组是 复制连接的后果，侧翼晚复制DNA 并可以解释与染色体变化有关的起源。 细胞的不朽 Neu原癌基因，人类的大鼠同源物 erb-B-2基因在两个大鼠乳腺癌组织中既未扩增也未过表达， 在体外被化学致癌物转化成肿瘤的细胞系。 编码人类胞质甲状腺激素基因的cDNA 结合蛋白（p58）通过原位杂交定位于15 q24 -25。 这种定位可以作为泰-萨克斯病的有用标记， 将允许评估染色体改变的影响， 对人类恶性肿瘤的影响