MOLECULAR MECHANISMS IN NEOPLASIA

该项目的总体目标是勘探和开发 逆转录病毒生物学作为一种手段，以更深入地了解肿瘤 变化 该模型系统的大部分拟议的工作是 重点是在鸡的法氏囊中诱导淋巴瘤 通过自然和基因工程逆转录病毒。 具体实验 旨在探索特定癌基因在诱导中的作用， 肿瘤前转化卵泡阶段和进展中 从转化的卵泡到更晚期的肿瘤。 录病毒 将制备携带c-myc和ChBlym-1基因的构建体，在 组织培养系统，并引入胚胎囊干细胞， 法氏囊移植技术。 癌前病变和 将对这种方法诱导的晚期肿瘤进行评价， 使用核酸和免疫学方法检测相关癌基因的表达 probes. 这些和其他原癌基因的正常表达将是 使用逆流离心法在法氏囊淋巴细胞中进行评价 分离淋巴细胞群的技术。 原位杂交将 用于放大物理分离技术获得的结果。 我们将寻找腔上囊生理性促有丝分裂刺激的证据， 淋巴细胞在发育过程中 除c-myc以外的原癌基因， 例如，c-fos通常被这个过程激活， 其他潜在的有趣的基因，将测试它们的影响， 法氏囊移植试验。 最后，将尝试 开发表达反义RNA的逆转录病毒构建体， 以选择性地抑制该体内系统中的癌基因表达。

The overall goal of this project is the exploration and exploitation of the biology of retroviruses as a means to a deeper understanding of neoplastic change. The model system on which the majority of the proposed work is focused is the induction of lymphomas in the bursa of Fabricius of chickens by natural and genetically engineered retroviruses. Specific experiments are directed at exploring the role of specific oncogenes in the induction of the pre-neoplastic transformed follicle stage and in the progression from transformed follicles to more advanced neoplasms. Retroviral constructs bearing c-myc and ChBlym-1 genes will be prepared, evaluated in tissue culture systems and introduced into embryonic bursal stem cells by the technique of bursal transplantation. Preneoplastic lesions and advanced neoplasms induced by this methodology will be evaluated for the expression of the relevant oncogenes using nucleic acid and immunologic probes. The normal expression of these and other proto-oncogenes will be evaluated in bursal lymphocytes using a counterflow centrifugation technique to separate lymphocyte populations. In situ hybridization will be used to amplify results obtained by the physical separation technique. Evidence will be sought of physiological mitogenic stimulation of bursal lymphocytes during development. Proto-oncogenes other than c-myc, for example, c-fos which are normally activated by this process, and a number of other potentially interesting genes, will be tested for their effects in the bursal transplantation assay. Finally, an attempt will be made to develop retroviral constructs expressing anti-sense RNA which might be used to selectively inhibit oncogene expression in this in vivo system.