Gerlach Bärbel, Harder Anna H, Hulsebos Theo J M, Leenstra Sieger, Slotman Berend J, Vandertop W Peter, Hartmann Karl-Axel, Sminia Peter
Department of Radiation Oncology, VU University Medical Center, Amsterdam, Netherlands.
Strahlenther Onkol. 2002 Sep;178(9):491-6. doi: 10.1007/s00066-002-0952-5.
Determination of in-vitro radiosensitivity and genetic alterations of cell cultures derived from human glioma biopsy tissue and established glioma cell lines.
Fresh brain tumor specimens of six patients were processed to early passage cell cultures. In addition the cell lines D 384 and Gli 6 were used. Cell cultures were irradiated with doses from 2 to 10 Gy. Following irradiation, cell survival was determined by clonogenic assay and survival curves were generated. The surviving fractions after 2 Gy (SF2) and 4 Gy (SF4) were used as radiosensitivity parameters. Genetic analysis included determination of the mutational and loss of heterozygosity (LOH) status of TP 53 (exons 5-8), the LOH 10- and epidermal growth factor receptor gene (EGFR) amplification status.
The SF2 and SF4 values ranged from 0.54 to 0.88 (mean: 0.70) and from 0.13 to 0.52 (mean: 0.32), respectively. Genetic alterations were found in the Gli 6 cell line and in two primary cell cultures. The genetic profile of Gli 6 showed LOH but no TP 53 mutation, complete LOH 10 and no EGFR amplification. The VU 15 cell culture showed TP 53 mutation but no LOH 10 or EGFR amplification, while VU 24 showed incomplete LOH 10, EGFR amplification and no TP 53 mutation. In the other four cell cultures and D 384 cell line no genetic alterations were diagnosed. Histopathological classification of glioblastoma multiforme and/or genetic alterations resulted in lower radiosensitivity.
In this small series of early passage glioma cell cultures low radiosensitivity and alterations in cell regulatory genes were seen. Further testing of biological behavior in larger series of patient-derived material is ongoing.
测定源自人胶质瘤活检组织和已建立的胶质瘤细胞系的细胞培养物的体外放射敏感性和基因改变。
对6例患者的新鲜脑肿瘤标本进行处理，建立早期传代细胞培养物。此外，还使用了D 384和Gli 6细胞系。细胞培养物接受2至10 Gy的剂量照射。照射后，通过克隆形成试验测定细胞存活率并生成存活曲线。将2 Gy（SF2）和4 Gy（SF4）后的存活分数用作放射敏感性参数。基因分析包括测定TP 53（外显子5 - 8）的突变和杂合性缺失（LOH）状态、10号染色体杂合性缺失（LOH 10）以及表皮生长因子受体基因（EGFR）扩增状态。
SF2和SF4值分别在0.54至0.88（平均值：0.70）和0.13至0.52（平均值：0.32）之间。在Gli 6细胞系和两种原代细胞培养物中发现了基因改变。Gli 6的基因图谱显示存在LOH但无TP 53突变，10号染色体完全杂合性缺失且无EGFR扩增。VU 15细胞培养物显示TP 53突变但无10号染色体杂合性缺失或EGFR扩增；而VU 24显示10号染色体不完全杂合性缺失、EGFR扩增且无TP 53突变。在其他四种细胞培养物和D 384细胞系中未诊断出基因改变。多形性胶质母细胞瘤的组织病理学分类和/或基因改变导致放射敏感性降低。
在这一小系列早期传代的胶质瘤细胞培养物中，观察到低放射敏感性和细胞调节基因的改变。正在对更多患者来源的材料进行更大规模的生物学行为进一步测试。
