CHROMOSOME ALTERATIONS AND PROTO-ONCOGENES TRANSPOSITION IN CARCINOGENESIS

细胞遗传学和分子遗传学相结合的方法将导致 识别具有显著影响的重要基因和基因改变 在癌症控制中的临床应用。虽然集成了 细胞基因组中的人乳头瘤病毒(HPV)DNA不会导致 病毒产生、HPV整合是宫颈癌的标志。 人乳头瘤病毒在染色体脆性部位和原癌基因附近的整合 我们的研究表明，这既是一种结构要求，也是一种 表型的生物选择。来自的几个集成站点 HPV永生化角质形成细胞系的高分辨率特征 病毒G-带和DNA相关DNA序列的原位定位 复制模式。HPV-16序列被定位在G-Light带上 表现出较晚的DNA复制。一种适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 被引入来研究癌症发生过程中的基因组变化。通过 荧光原位杂交(FISH)和数字成像 在C4-1宫颈癌中定位了HPV-18整合位点。一个DNase I- 在病毒DNA的3kb范围内发现了过敏性位点。在……里面 此外，与肿瘤相比，整合区域的甲基化程度较低 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呈现 病毒整合可获得的特定基因组位置。鱼曾经是 优化并用于检测完整核上的核RNA。在一个 单拷贝整合Epstein-Barr基因的Burkitt淋巴瘤细胞系 病毒，一种反映RNA位置的特异杂交信号 转录直接显示在完整的细胞核上。把鱼捞进去 与增强的数字处理相结合，使检测和 相对较小的单拷贝基因的作图。一场变革 Est基因和一个胆囊素-胃泌素受体基因被定位在 人类10号染色体(p11.2)和11号染色体(p15.5)。这两个基因都可能 参与人类癌症的发生。

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.