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: 3752668
• 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/3752668
• 关键词: cervix neoplasms; chromosome translocation; digital imaging; drug resistance; gene expression; genetic mapping; genetic markers; human papillomavirus; human tissue; in situ hybridization; laboratory rat; neoplasm /cancer genetics; neoplastic cell; neoplastic process; neoplastic transformation; nucleic acid sequence; oncogenes; polymerase chain reaction; protooncogene; transfection; tumor suppressor genes; viral carcinogenesis; virus genetics

Fluorescence in situ hybridization (FISH) and digital-imaging analysis significantly enhance the identification of complex or cryptic chromosomal changes in cancer cells. With multiprobe FISH, nonrandom deletions of chromosomes 1, 3, 9, and 11 have been identified in human papillomavirus (HPV)-negative cervical carcinomas--most likely, delineating regions of tumor-suppressor genes. Cumulative chromosomal changes associated with the process of neoplastic transformation of human exocervical cells immortalized by HPV-16 DNA and converted to tumorigenicity by Ha-ras cotransfection was demonstrated by FISH and multicolor detection. The progression towards tumorigenicity was associated with chromosome 1, 3, 9, and 17 anomalies which commonly occur in cervical carcinomas and demonstrate the relevance of in vitro models for studying cervical neoplasia. FISH also substantially increases the resolution of mapping cancer-related genes in mammalian chromosomes. Among several new genes mapped, two histone genes, H2A.Z and H2A.X, were localized to chromosomes 4q24 and 11q23.2-23.3, respectively. The latter site is a highly recombinogenic region in the human genome. A putative multidrug resistance (MDR) enhancerlike element was located 10 kb upstream to the proximal MDR1-P1 promoter on chromosome 7. FISH analysis revealed that this enhancerlike element is on chromosome 20q13.1 and is distinct from the MDR locus on chromosome 7. Using FISH with probes directly biotinylated by polymerase chain reaction, single-copy genes of 200-400 bp were localized on metaphase chromosomes. Reciprocal chromosomal translocations, amplification, direct visualization of multiple members of a gene family, or order of genes within a genomic site were demonstrated using this approach. This new development has unlimited potential for the simultaneous discovery and mapping of new genes, for identification of markers associated with neoplasia and other disorders, as well as for position-cloning strategy.

荧光原位杂交（FISH）和数字成像分析 显著增强对复杂或隐蔽的 癌细胞的染色体变化 使用多探针FISH，非随机 已经在人类中鉴定了染色体1、3、9和11的缺失 乳头瘤病毒阴性的宫颈癌--很可能， 描绘肿瘤抑制基因的区域。 累积染色体 与人类肿瘤转化过程相关的变化 HPV-16 DNA永生化的外生宫颈癌细胞， FISH证实Ha-ras共转染的致瘤性， 探测器。 向致瘤性的进展是 与染色体1、3、9和17异常相关， 并证明了体外模型的相关性 用于研究宫颈肿瘤 FISH还大大增加了 在哺乳动物染色体中绘制癌症相关基因的分辨率。 在几个新定位的基因中，两个组蛋白基因H2A.Z和H2A.X被发现， 分别位于染色体4 q24和11q23.2-23.3。 后者 位点是人类基因组中高度重组的区域。 推定的 多药耐药（MDR）增强子样元件位于10 kb 位于7号染色体上的近端MDR 1-P1启动子上游。 FISH分析 揭示了这种增强子样元件位于染色体20q13.1上， 与7号染色体上的MDR基因座不同。 使用探针进行FISH 通过聚合酶链反应直接生物素化， 200-400 bp定位于中期染色体。 互惠 染色体易位，扩增，直接可视化 基因家族的多个成员，或基因组中基因的顺序 网站使用这种方法进行了演示。 这一新的发展 无限的潜力，同时发现和映射新的 基因，用于鉴定与瘤形成和其他 疾病以及位置克隆策略。