GENETIC ALTERATIONS IN PROGRESSION OF HUMAN MELANOMA

问题是：各种显性行为、隐性行为或 不同基因中的显性-负性突变是最常见的 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上取得的进展，例如。结直肠癌， 肺癌和乳腺癌尤其令人印象深刻。 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，但 临床材料和已建立的细胞系的可用性，以及巨大的 它被研究的程度，没有显性的癌基因-与 碱性成纤维细胞生长因子基因或抑制子异位表达的可能例外 类型抑癌基因-在人类恶性肿瘤中一直被发现 黑色素瘤。 假设和目标：染色体转移实验和逐步 人类黑色素瘤的进化发展强烈地涉及 一些基因参与了这一疾病的发生和发展 疾病，其中至少一个或多个看起来是隐性肿瘤 基于最近的染色体转移实验的抑制基因。它是 假设其中一些基因的身份可以通过 逆转录病毒载体插入突变(前病毒)技术 标记)和分子克隆。 实验方法：两个人细胞系，一个是放射状生长期(RGP) 已知的黑色素瘤在裸鼠体内不会致癌，另一种是早期 垂直生长期(VGP)黑色素瘤已知致癌-但 转移性-无能力-将感染一种两性营养包装 含有显性药物可选择遗传标记的小鼠逆转录病毒。 病毒感染成功的细胞将被注射到裸鼠体内 使用原位(皮下)移植程序。由此产生的 RGP来源的原发肿瘤或VGP来源的转移将被分析为 常见的前病毒整合部位。宿主基因组DNA侧翼这样 整合地点将被克隆，作为识别假设的手段 导致黑色素瘤进展的显性或隐性基因。

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.