Is 24-color FISH detection of in-vitro radiation-induced chromosomal aberrations suited to determine individual intrinsic radiosensitivity?
Is 24-color FISH detection of in-vitro radiation-induced chromosomal aberrations suited to determine individual intrinsic radiosensitivity?
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DOI：
10.1007/s00066-002-0904-0
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发表时间：
2002-04-01
影响因子：
3.1
通讯作者：
Wendt, TG
中科院分区：
文献类型：
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作者：
Kuechler, A;Neubauer, S;Wendt, TG
背景：在放射肿瘤学中，可靠地测定个体患者的固有放射敏感性是一个迫切的需求。染色体畸变是以前暴露于电离辐射的敏感指标。以前的分子细胞遗传学研究表明，这种畸变相当于内在的辐射敏感性，可以检测荧光原位杂交（FISH）技术使用全染色体彩绘（WCP）探针。然而，到目前为止，只有一个到三个随机选择的wcp探针已应用于这种方法。作为一个随机分布的染色体重排沿着的染色体是到现在为止仍然有争议的权力，24色FISH的方法应阐明在本study.Methods和材料：淋巴细胞来源于淋巴母细胞样细胞系的一名患者Nijmegen断裂综合征（NBS纯合子）和两个NBS杂合子和外周血淋巴细胞的两个对照进行了分析。每例患者/对照的样本。在培养前用0.0戈伊、0.7戈伊或2.0戈伊进行体外照射。染色体畸变进行了详细的分析和量化的24色FISH作为个人内在的radiosensitivity.Results表达的手段，24色FISH分析共1,674中期。经体外照射后，对照组细胞的畸变率为21%（0.7戈伊）或57%（2.0戈伊），杂合子组细胞的畸变率为15%（0.7戈伊）或53%（2.0戈伊），纯合子组细胞的畸变率为54%（0.7戈伊）或79%（2.0戈伊）。在NBS患者中观察到最高的平均有丝分裂断裂率[B/M]（0.7 Gy：1.80 B/M，2.0 Gy：4.03 B/M）和复杂染色体重排率[CCR]（0.7戈伊：0.20 CCR/M，2.0戈伊：0.47 CCR/M）。此外，照射后不同畸变类型的比例显示，NBS纯合子中CCR率明显增加，双着丝粒减少。为了更全面地了解辐射诱导的畸变，并检测和量化遗传决定的内在辐射敏感性，使用全人类染色体涂染探针的24色FISH方法已成功地应用于细胞遗传学制备淋巴细胞。对照和NBS杂合子与NBS纯合子受试者明显不同。
Background: Reliable determination of intrinsic radiosensitivity in individual patients is a serious need in radiation oncology. Chromosomal aberrations are sensitive indicators of a previous exposure to ionizing irradiation. Former molecular cytogenetic studies showed that such aberrations as an equivalent of intrinsic radiosensitivity can be detected by fluorescence in-situ hybridization (FISH) techniques using whole chromosome painting (wcp) probes. However, only one up to three randomly chosen wcp probes have been applied for such approaches until now. As a random distribution of chromosomal rearrangements along the chromosomes is up to now still controversial, the power of the 24-color FISH approach should be elucidated in the present study.Methods and Material: Lymphocytes derived from lymphoblastoid cell Lines of one patient with Nijmegen breakage syndrome (NBS homozygote) and of two NBS heterozygotes and peripheral blood lymphocytes of two controls were analyzed. Samples of each patient/control. were irradiated in vitro with 0.0 Gy, 0.7 Gy or 2.0 Gy prior to cultivation. Chromosomal aberrations were analyzed in detail and quantified by means of 24-color FISH as an expression of the individual intrinsic radiosensitivity.Results; 24-color FISH analyses were done in a total of 1,674 metaphases. After in-vitro irradiation, 21% (0.7 Gy) or 57% (2.0 Gy) of the controls' cells, 15% (0.7 Gy) or 53% (2.0 Gy) of the heterozygotes' cells and 54% (0.7 Gy) or 79% (2.0 Gy) of the homozygote's cells contained aberrations. The highest average rates of breaks per mitosis [B/M] (0.7 Gy: 1.80 B/M, 2.0 Gy: 4.03 B/M) and complex chromosomal rearrangements [CCR] (0.7 Gy: 0.20 CCR/M, 2.0 Gy: 0.47 CCR/M) were observed in the NBS patient. Moreover, the proportion of different aberration types after irradiation showed a distinct increase in the rate of CCR combined with a decrease in dicentrics in the NBS homozygote.Conclusion: To come to a more complete picture of radiation-induced aberrations and to detect and quantify genetically determined intrinsic radiosensitivity, a 24-color FISH approach using all human chromosome painting probes has been successfully applied on cytogenetic preparation lymphocytes. The controls and NBS heterozygotes were clearly distinguished from the NBS homozygote subject.