Tumor Suppressors in Lymphocytes

描述（由申请人提供）：正常、受控细胞生长所必需的基因称为肿瘤抑制基因，这些基因的失活可导致肿瘤形成。大多数已知的肿瘤抑制因子是通过遗传易感性为癌症的生物体的遗传图谱定位的。然而，家族性癌症易感性仅导致约10%的人类癌症，并且涉及非家族性癌症的肿瘤抑制因子的鉴定较慢。该提案的目标是定义一组肿瘤抑制因子，当删除时有助于小鼠淋巴细胞肿瘤的形成;以及它们与原癌基因的相互作用，尽可能完整。这将通过逆转录病毒插入诱变与化学诱变相结合来尝试，以在给定小鼠的细胞中使两个等位基因突变。通过逆转录病毒对化学诱变雄性小鼠的后代进行插入诱变，诱导淋巴细胞来源的肿瘤。病毒基因组破坏潜在的抑制基因，导致其失活，同时产生用于鉴定插入位点的标记。标记的癌症基因将通过基于高通量PCR的技术进行克隆和测序。随后，将进一步表征它们的子集，并确定它们与其他癌基因（共突变）合作的性质。

DESCRIPTION (provided by applicant): The genes that are necessary for normal, controlled cell growth are called tumor suppressor genes, and inactivation of these genes can lead to tumor formation. The majority of known tumor suppressors were located by the genetic mapping of organisms with an inherited predisposition for cancer. However, familial predisposition to cancer is responsible for only approximately 10% of human cancers and identification of tumor suppressors involved in non-familial cancers has been slower. The goal of this proposal is to define the set of tumor suppressors that when deleted contribute to the formation of lymphocyte tumors in mice; and their interactions with protooncogenes, as completely as possible. This will be attempted by retroviral insertional mutagenesis, combined with chemical mutagenesis, to inactivate both alleles in cells of a given mouse. The offspring of chemically mutagenized male mice are subjected to insertional mutagenesis by retrovirus that induces tumors of lymphocyte origin. The viral genome disrupts potential suppressor genes, leading to their inactivation, and at the same time creates a marker for identifying the insertion loci. The tagged cancer genes will be cloned and sequenced by a high throughput PCR based technique. Subsequently, a subset of them will be further characterized and the nature of their cooperation with other oncogenes (co-mutations) will be determined.