IN VITRO CULTURE OF RADIATION INDUCED PRELEUKEMIC CELL

致白血病剂量的阿司匹林对骨髓的初始改变 将对辐射进行检查。将采用多种组织培养方法 用来繁殖和丰富那些可以诱导 白血病在过继转移实验中。这些改变的目标细胞将 检测其表达的生态亲和性和重组病毒 指示剂共培养试验。还将对这些细胞进行检查 用Southern分析内源性前病毒遗传元件的变化 他们的DNA的限制性内切酶。同样，已知的一般位置 癌基因将在限制性内切酶中进行分析，它们的表达将 用定量斑点杂交法进行测定。 其他转化相关蛋白和生长因子的表达 将由新陈代谢合成试验确定，使用 免疫沉淀/-聚丙烯酰胺凝胶分析。生长发育的生物测定 将利用白介素2和白介素3等因素来确定可能的 这些生长因子和其他生长因子的靶细胞表达的变化。 骨髓基质支持正常生长的能力，并改变了 辐射骨髓基质的生长因子也将在 部分和完全白血病诱导方案。 这样的研究将导致确定义务与巧合 骨髓的改变，导致小鼠白血病的产生 电离辐射。

The initial alterations induced in bone marrow by leukemogenic doses of radiation will be examined. A variety of tissue culture methods will be utilized to propagate and enrich those target cells that can induce leukemia in adoptive transfer experiments. These altered target cells will be examined for their expression of ecotropic and recombinant virus in indicator co-cultivation assays. The cells will also be examined for alterations in endogenous proviral genetic elements by Southern analysis of restriction digests of their DNA. Similarly, the general location of known oncogenes will be analyzed in restriction digests and their expression will be measured by a quantitative dot blot hybridization assay. Expression of other transformation associated proteins and growth factors will be determined by metabolic synthesis assays using immunoprecipitation/-polyacrylamide gel analysis. Bioassays for growth factors such as interleukin 2 and 3 will be utilized to determine possible alterations in target cell expression of these and other growth factors. Marrow stromal capacity to support normal growth, and altered production of growth factors by radiated marrow stroma will also be examined during partial and complete leukemia induction protocols. Such studies will lead to a determination of obligate vs. coincidental alterations in bone marrow, resulting in generation of murine leukemia by ionizing radiation.