Zemanová Z, Kramar F, Babická L, Ransdorfová S, Melichercíková J, Hrabal P, Kozler P, Michalová K
Centre of Oncocytogenetics, Institute of Clinical Biochemistry and Laboratory Diagnostics, 1st Faculty of Medicine, Charles University, and General Teaching Hospital, Prague, Czech Republic.
Folia Biol (Praha). 2006;52(3):71-8.
In oligodendroglial brain tumours, losses of chromosomal material of the short arm of chromosome 1 and long arm of chromosome 19 have been shown to predict responsiveness to chemotherapy and prolonged patients' survival. Therefore, the correct diagnosis of these genetic alterations in tumours of oligodendroglial origin is particularly important. To detect deletions of 1p36 and/or 19q13.3 in oligodendroglial cells we used dual-colour I-FISH with locus-specific DNA probes. I-FISH was performed on isolated whole cell nuclei, prepared from fresh non-fixed tumour tissue samples resuspended in media and processed using a standard cytogenetic procedure, thus bypassing the problem of nuclear truncation. We examined 16 patients with histologically proved oligodendrogliomas (5x oligodendroglioma, 9x anaplastic oligodendroglioma, 2x anaplastic oligoastrocytoma). The results of molecular cytogenetic analyses were correlated with morphological and clinical findings. Molecular cytogenetic analyses were successful in 15 patients and, due to a non-adequate tissue specimen, were uninformative in one patient only. Combined deletions 1p36/19q13 were proved in 13 patients. However, in six of them additional genetic alterations typical for high-grade astrocytoma were found, which could have negative influence on the prognosis. One patient had isolated deletion of 1p36 and another had a normal genetic pattern without any chromosomal alterations. In summary, I-FISH on isolated cell nuclei is a powerful tool for detecting chromosomal aberrations in tumour cells. A systematic molecular cytogenetic analysis may advance diagnosis, prognostic stratification, and targeted treatment of patients with brain tumours.
在少突胶质细胞瘤中,1号染色体短臂和19号染色体长臂的染色体物质缺失已被证明可预测对化疗的反应及延长患者生存期。因此,正确诊断少突胶质细胞起源肿瘤中的这些基因改变尤为重要。为检测少突胶质细胞中1p36和/或19q13.3的缺失,我们使用了针对特定基因座的DNA探针进行双色原位荧光杂交(I-FISH)。I-FISH是在从新鲜未固定的肿瘤组织样本中分离出的全细胞核上进行的,这些样本重悬于培养基中,并采用标准细胞遗传学程序进行处理,从而避免了核截断问题。我们检查了16例经组织学证实的少突胶质细胞瘤患者(5例少突胶质细胞瘤、9例间变性少突胶质细胞瘤、2例间变性少突星形细胞瘤)。分子细胞遗传学分析结果与形态学和临床发现相关。分子细胞遗传学分析在15例患者中成功进行,仅1例患者因组织标本不足而未获得有效信息。13例患者被证实存在1p36/19q13联合缺失。然而,其中6例还发现了高级别星形细胞瘤典型的其他基因改变,这可能对预后产生负面影响。1例患者存在1p36孤立缺失,另1例患者基因模式正常,无任何染色体改变。总之,对分离细胞核进行I-FISH是检测肿瘤细胞染色体畸变的有力工具。系统的分子细胞遗传学分析可能会推动脑肿瘤患者的诊断、预后分层和靶向治疗。