GROWTH AND RADIATION RESPONSE OF AN IN VITRO TUMOR MODEL

这项研究的目的是调查细胞状态的变化， 影响细胞的辐射敏感性，并可由环境诱导 肿瘤中所见的那种剥夺，即氧气/营养剥夺或 剥夺后再灌流。对影响的变化进行分析 敏感性分为三个部分：a)定量评估 细胞的DNA水平和辐射反应的变化 修饰的DNA；b)基本的、动态的演示和测量 细胞活动的变化(如分化、药物摄取)；c) 检测指示先前细胞的固有亚细胞标志物 剥夺和改变了辐射敏感性的修饰表型。 三明治系统(体外细胞培养)将用于实验 含有营养物质和代谢物的类肿瘤细胞群模型 由于距离血管形成的距离而产生的梯度。在三明治中，如 在肿瘤中，在环境条件下施加的梯度在 细胞表型和细胞放射反应。许多检测到的 要使用的分析方法利用了在现场进行检测的能力 单细胞水平上的测量，并且不会丢失关于 种群内细胞的空间位置。它的几何形状 三明治制度诱导种群异质性在空间上发展 有条理的方式。因此，理清因果关系就成了 是可以实现的。不同细胞亚群间的相互作用 字符存在，但由于单层性质，完整的细胞访问和 保持可见性，允许对两个单独单元进行评估 敏感度和整体反应。 拟议的工作包括实验和理论两个部分。首字母 研究展示了由我们的梯度方法揭示的新的生物学结果 对于剥夺诱导的现象(例如，DNA含量和 亚核分布，诱导分化，增加 线粒体膜电位)。

The aim of this study is to investigate alterations in cell state which affect cellular radiosensitivity and are inducible by environmental deprivation of the kind seen in tumors, i.e. oxygen/nutrient deprivation or reperfusion following deprivation. The analysis of changes affecting sensitivity is divided into three parts: a) quantitative assessment of changes at the DNA level and the radioresponse of the cells exhibiting modified DNA; b) demonstration and measurement of fundamental, dynamic alterations in cellular activities (e.g. differentiation, drug uptake); c) detection of intrinsic subcellular markers indicative of previous cellular deprivation and of a modified phenotype with altered radiosensitivity. The sandwich system (in vitro cell-culture) will be used to experimentally model tumor-like populations of cells having nutrient and metabolite gradients arising due to distance from vascularization. In sandwiches, as in tumors, imposed gradients in environmental condition induce gradients in cellular phenotype and cellular radioresponse. Many of the detection assays to be used take advantage of the ability to make in situ measurements on the single cell level and not lose information on the spatial location of cells within the population. The geometry of the sandwich system induces population heterogeneity to develop in a spacially organized manner. Thus, sorting out of cause and effect becomes achievable. Interactions between subpopulations of cells of differing character exist, yet due to the monolayer nature, complete cell access and visibility are maintained, permitting assessment of both individual cell sensitivity and whole population response. The proposed work has experimental and theoretical components. Initial studies present novel biological results revealed by our gradient approach to deprivation-induced phenomena (e.g. alterations in DNA content and sub-nuclear distribution, induction of differentiation, increased mitochondrial membrane potential).