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: 3774829
• 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/3774829
• 关键词: Burkitt's lymphoma; Epstein Barr virus; Papillomavirus; carcinoma; cervix neoplasms; chromosome translocation; digital imaging; gene expression; gene induction /repression; gene rearrangement; genetic mapping; genetic transcription; human herpesvirus 6; human tissue; in situ hybridization; laboratory rat; neoplasm /cancer genetics; neoplastic transformation; nucleic acid sequence; oncogenes; protooncogene; transfection; viral carcinogenesis; virus genetics

Combined cytogenetic and molecular genetic approaches will lead to the recognition of important genes and gene alterations that have significant clinical application in the control of cancer. Although integration of human papillomavirus (HPV) DNA in the cellular genome does not result in virus production, HPV integration is a hallmark of cervical carcinoma. HPV integration at chromosome fragile sites and near proto-oncogenes demonstrated by our studies suggests both a structural requirement and a biological selection of the phenotype. Several integration sites from HPV-immortalized keratinocyte lines were characterized by high-resolution in situ localization of viral DNA sequences relative to G-band and DNA replication patterns. HPV-16 sequences were mapped at G-light bands exhibiting late DNA replication. A system for digital imaging suitable for detection and analysis of a variety of immunofluorescent cytological reactions and for detection of single-copy genes or virus copies was introduced to investigate genomic alterations in carcinogenesis. By fluorescent in situ hybridization (FISH) and digital imaging, a single HPV-18 integration site was mapped in C4-1 cervical carcinoma. A DNase I- hypersensitive site was identified within 3 kb from the viral DNA. In addition, the integration region was undermethylated compared to tumor cell lines of other origins. Molecular and cytological evidence demonstrate that structural and functional chromatin features render specific genomic sites accessible to viral integration. FISH was optimized and used for detection of nuclear RNA on intact nuclei. In a Burkitt lymphoma cell line with a single-copy integrated Epstein-Barr virus, a specific hybridization signal reflecting the site of RNA transcription was directly visualized on intact nuclei. FISH in conjunction with enhanced digital processing allowed the detection and mapping of single-copy genes of relatively small size. A transforming gene (est) and a cholecystokinin-gastrin receptor gene were mapped in human chromosomes 10 (p11.2) and 11 (p15.5), respectively. Both genes may be involved in human carcinogenesis.

结合细胞遗传学和分子遗传学方法将导致 识别重要的基因和基因改变， 在癌症控制中的临床应用。 虽然整合 细胞基因组中的人乳头瘤病毒（HPV）DNA不会导致 HPV整合是宫颈癌的标志。 HPV在染色体脆性位点和原癌基因附近的整合 我们的研究表明，既有结构要求， 表型的生物选择。 来自的多个集成站点 HPV永生化的角质形成细胞系的特征在于高分辨率 病毒DNA序列相对于G带和DNA的原位定位 复制模式 HPV-16序列定位于G-光带 表现出晚期DNA复制。 一种用于数字成像的系统， 用于检测和分析各种免疫荧光细胞学 反应和单拷贝基因或病毒拷贝的检测是 研究致癌过程中的基因组改变。 通过 荧光原位杂交（FISH）和数字成像，单一 HPV-18整合位点在C4-1宫颈癌中的定位。DNA酶I- 在病毒DNA的3 kb范围内鉴定出高敏感位点。 在 此外，与肿瘤相比，整合区甲基化不足 其他来源的细胞系。 分子和细胞学证据 证明结构和功能染色质特征使 病毒整合的特定基因组位点。 鱼 优化并用于检测完整核上的核RNA。 中 具有单拷贝整合的Epstein-Barr的Burkitt淋巴瘤细胞系 病毒，反映RNA位点的特异性杂交信号 在完整的细胞核上直接观察转录。 鱼 与增强的数字处理相结合， 相对较小的单拷贝基因的定位。 变换 基因（est）和胆囊收缩素-胃泌素受体基因的定位， 人染色体10（p11.2）和11（p15.5）。 这两种基因可能 与人类致癌有关。