MOLECULAR MECHANISMS IN NEOPLASIA

该项目的总体目标是勘探和开发 逆转录病毒生物学是加深对肿瘤认识的一种手段 变化。建议的大部分工作所基于的模型系统 焦点是鸡法氏囊淋巴瘤的诱发 通过自然和基因工程的逆转录病毒。具体实验 旨在探索特定癌基因在诱导中的作用 肿瘤前转化的卵泡期及其进展 从转化的卵泡到更高级的肿瘤。逆转录病毒 含有c-myc和ChBlym-1基因的构建体将在 组织培养体系并通过以下方法导入胚胎法氏囊干细胞 法氏囊移植技术。癌前病变和 通过这种方法诱发的晚期肿瘤将被评估为 用核酸和免疫学方法研究相关癌基因的表达 探测器。这些和其他原癌基因的正常表达将是 逆流离心法在法氏囊淋巴细胞中的应用 分离淋巴细胞群的技术。原位杂交将 用于放大通过物理分离技术获得的结果。 将寻找法氏囊生理性有丝分裂刺激的证据 发育过程中的淋巴细胞。C-myc以外的原癌基因，例如 例如，通常由该进程激活的c-fos，以及一个数字 其他可能令人感兴趣的基因，将在 法氏囊移植试验。最后，我们将尝试 发展表达反义RNA的逆转录病毒载体，可能用于 在这个体内系统中选择性地抑制癌基因的表达。

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.