Walsh J W, Zimmer S G, Oeltgen J, Markesbery W R
Neurosurgery. 1986 Aug;19(2):185-200. doi: 10.1227/00006123-198608000-00003.
This report presents an experimental model for study of the cellular and molecular biology of invasiveness in tumors. It uses SV40 virus for the production of primary intracranial tumors that are invasive for normal brain and vary markedly and predictably in this invasiveness. Cell cultures of dissociated 1- to 2-day-old Syrian hamster cerebral cortex (Cx), brain stem (Bs), cerebellar hemisphere (Cbh), and cerebellar vermis (Cbv) were transformed with SV40 virus and inoculated intracerebrally into newborn hamsters. All 368 animals that developed intracranial tumors were killed, and tumor was taken for histological and immunofluorescence studies, assessment of extent of invasiveness, and preparation of cell cultures from which cells were cloned by dilution plating or growth in soft agar. A few hamsters were perfused with glutaraldehyde for studies of tumor ultrastructure. All cloned and uncloned tumor cells were reinoculated to produce second- and third-passage tumors. Characteristic differences in morphology and growth rate were observed between normal astrocytes derived from each brain region, and these phenotypic differences were retained after virus transformation and tumor production. Cloned and uncloned Cx cell-derived tumors of second and third passage diffusely invaded adjacent normal brain, although those of first passage invaded only slightly. Except for extracerebral spread, these tumors resembled human astrocytic series tumors. Bs and some Cbh cell-derived tumors were also astrocytic but more undifferentiated and only slightly invasive; Cbv and other Cbh cell-derived tumors were sarcomatous and only extended along perivascular spaces or were not invasive at all. The tumor cells contained glial fibrillary acidic protein and SV40 T-antigen. These results suggest that astrocytes from different brain regions vary in genomic stability and support the theory that differences in invasiveness reflect the development of heterogeneity and subsequent selection of more aggressive subpopulations of cells.
本报告介绍了一种用于研究肿瘤侵袭性细胞生物学和分子生物学的实验模型。它利用SV40病毒产生原发性颅内肿瘤，这些肿瘤可侵袭正常脑实质，且在侵袭性方面有显著且可预测的差异。将1至2日龄叙利亚仓鼠的大脑皮层（Cx）、脑干（Bs）、小脑半球（Cbh）和小脑蚓部（Cbv）解离后的细胞培养物用SV40病毒转化，并脑内接种到新生仓鼠体内。所有368只发生颅内肿瘤的动物均被处死，取出肿瘤进行组织学和免疫荧光研究、侵袭程度评估，并制备细胞培养物，通过稀释铺板或软琼脂培养进行细胞克隆。少数仓鼠用戊二醛灌注以研究肿瘤超微结构。所有克隆和未克隆的肿瘤细胞均再次接种以产生第二代和第三代传代肿瘤。观察到源自每个脑区的正常星形胶质细胞在形态和生长速率上存在特征性差异，并且这些表型差异在病毒转化和肿瘤形成后得以保留。第二代和第三代传代的克隆和未克隆的Cx细胞源性肿瘤广泛侵袭相邻的正常脑实质，而第一代传代肿瘤仅略有侵袭。除脑外扩散外，这些肿瘤类似于人类星形细胞系列肿瘤。Bs和一些Cbh细胞源性肿瘤也是星形细胞性的，但分化程度更低且侵袭性较弱；Cbv和其他Cbh细胞源性肿瘤为肉瘤性，仅沿血管周围间隙扩展或根本无侵袭性。肿瘤细胞含有胶质纤维酸性蛋白和SV40 T抗原。这些结果表明，来自不同脑区的星形胶质细胞在基因组稳定性方面存在差异，并支持侵袭性差异反映异质性发展以及随后更具侵袭性的细胞亚群选择的理论。
