GENETIC ALTERATIONS IN PROGRESSION OF HUMAN MELANOMA

人类黑色素瘤进展过程中的基因改变

• 批准号: 3423587
• 负责人: YAACOV BEN DAVID
• 金额: $ 4万
• 依托单位: SUNNYBROOK & WOMEN'S COLL HLTH SCIS CTR
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-01 至 1993-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3423587
• 关键词: Retroviridae; athymic mouse; genetic markers; human tissue; melanoma; metastasis; molecular cloning; neoplasm /cancer genetics; neoplasm /cancer transplantation; neoplastic cell; neoplastic growth; phenotype; provirus; transfection; transposon /insertion element; tumor suppressor genes; viral carcinogenesis

THE PROBLEM: The identity of various dominantly acting, recessive, or dominant-negative mutations in different genes represents one of the most significant advances in our understanding of the origins of human cancer. The strides made in some forms of human cancer, eg. colorectal carcinoma, lung carcinomas and, breast cancer, have been particularly impressive. Malignant melanoma stands out as a conspicuous exception against this background of successes. Despite its rapid rise in incidence, the availability of clinical material and established cell lines, and the great extent to which it is being studied, no dominant oncogene - with the possible exception of ectopic expression of the bFGF gene, or suppressor type anti-oncogene - has been consistently noted in human malignant melanoma. HYPOTHESES and OBJECTIVES: Chromosome transfer experiments and the stepwise evolutionary development of human melanoma strongly implicate the involvement of a number of genes in the development and progression of this disease, at least one or more of which appear to be recessive tumor suppressor genes based on recent chromosome transfer experiments. It is hypothesized that the identity of some of these genes can be uncovered by the technique of retrovirus vector insertional mutagenesis (provirus tagging) and molecular cloning. EXPERIMENTAL APPROACH: Two human cell lines, one a radial growth phase (RGP melanoma known to be non-tumorigenic in nude mice, the other an early-stage vertical growth phase (VGP) melanoma known to be tumorigenic - but metastatically-incompetent - will be infected with an amphotrophic packaged murine retrovirus containing a dominant drug selectable genetic marker. Successfully viral infected cells will be injected into athymic nude mice using orthotopic (subdermal) transplantation procedures. Resultant RGP-derived primary tumors or VGP derived metastases will be analyzed for sites of common proviral integration. Host genomic DNA flanking such integration sites will be cloned as a means of identifying putative dominant or recessive genes which contribute to melanoma progression.

问题：各种显性行为，隐性行为， 不同基因中的显性负突变代表了 在我们对人类癌症起源的理解上取得了重大进展。 在某些形式的人类癌症方面取得的进展，例如。结肠直肠癌， 尤其是肺癌和乳腺癌， 恶性黑色素瘤是一个明显的例外 成功的背景。 尽管其发病率迅速上升， 临床材料和已建立的细胞系的可用性，以及 它正在研究的程度，没有显性癌基因-与 bFGF基因或抑制基因异位表达的可能例外 型抑癌基因-一直注意到在人类恶性肿瘤 黑素瘤 假设和假设：染色体转移实验和逐步 人类黑色素瘤的进化发展强烈暗示了 许多基因参与了这种疾病的发展和进展， 疾病，其中至少一个或多个似乎是隐性肿瘤 抑制基因的基础上最近的染色体转移实验。 是 假设这些基因中的一些基因的身份可以通过 逆转录病毒载体插入突变（provirus 标记）和分子克隆。 实验方法：两种人细胞系，一种处于放射状生长期（RGP 一种是已知在裸鼠中无致瘤性的黑色素瘤，另一种是早期黑色素瘤， 垂直生长期（VGP）黑色素瘤已知是致瘤性的，但 转移性无能-将感染一种包装的 含有显性药物选择性遗传标记的鼠逆转录病毒。 将成功感染病毒的细胞注射到无胸腺裸鼠中 使用原位（皮下）移植程序。 所得 将分析RGP来源的原发性肿瘤或VGP来源的转移， 共同前病毒整合的位点。 宿主基因组DNA侧接这样的 将克隆整合位点作为鉴定推定的 显性或隐性基因导致黑色素瘤进展。