Janka M, Fischer U, Tonn J C, Kerkau S, Roosen K, Meese E
Department of Neurosurgery, University of Würzburg, Saarland, Germany.
Anticancer Res. 1996 Sep-Oct;16(5A):2601-6.
Tumor fragment spheroids (TFS) represent an organotypic in vitro model with preserved cytoarchitecture and matrix components of the native tumor in situ. In order to determine whether DNA amplifications within gliomas remain stable in spheroid culture, tumor fragment spheroids were established from 15 human gliomas including 14 glioblastoma and one anaplastic astrocytoma. Native tumor tissue, monolayers as well as TFS were evaluated for DNA amplification using reverse chromosome painting (RCP). A modified protocol for DNA isolation from TFS was established. Amplifications in the original tumor tissue were found on chromosomes 12q13-15, tel, 4q12-13 and 11p12-13, an amplification on 11p12-13 is reported for the first time. By RCP we could demonstrate that amplified domains on 12q13-15 and 4q12-13 in three tumors were maintained in TFS whereas the amplification on 11p12-13 could not be confirmed in TFS. In monolayer cultures, all amplifications which were detected in primary culture were lost until passage 5. The results of this first comparative study of DNA amplification in glioma by analyzing native tumor tissue and tumor fragment spheroids enables us to conclude that TFS seems to be a promising in vitro model for the study of DNA amplification under cell culture conditions.
肿瘤碎片球体（TFS）代表一种体外器官型模型，其保留了原位天然肿瘤的细胞结构和基质成分。为了确定胶质瘤内的DNA扩增在球体培养中是否保持稳定，从15例人类胶质瘤（包括14例胶质母细胞瘤和1例间变性星形细胞瘤）中建立了肿瘤碎片球体。使用反向染色体描绘（RCP）对天然肿瘤组织、单层细胞以及TFS进行DNA扩增评估。建立了一种从TFS中分离DNA的改良方案。在原始肿瘤组织中发现12q13 - 15、端粒、4q12 - 13和11p12 - 13染色体存在扩增，其中11p12 - 13的扩增首次报道。通过RCP我们可以证明，三个肿瘤中12q13 - 15和4q12 - 13上的扩增区域在TFS中得以维持，而11p12 - 13上的扩增在TFS中未得到证实。在单层培养中，原代培养中检测到的所有扩增在传代至第5代时均消失。通过分析天然肿瘤组织和肿瘤碎片球体对胶质瘤中DNA扩增进行的首次比较研究结果使我们能够得出结论，TFS似乎是一种用于在细胞培养条件下研究DNA扩增的有前景的体外模型。
