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The Effect of X-Ray and Heavy Ions Radiations on Chemotherapy Refractory Tumor Cells

机译：X射线和重离子辐射对化疗难治性肿瘤细胞的影响

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Purpose The purpose of this study is to link both numeric and structural chromosomal aberrations to the effectiveness of radiotherapy in chemotherapy refractory tumor cells. Materials and methods Neuroblastoma (LAN-1) and 79HF6 glioblastoma cells derived from patients and their chemoresistant sublines were artificially cultured as neurospheres and irradiated by X-rays and heavy ions sources. All the cell lines were irradiated by Carbon-SIS with LET of 100?keV/μm. However, 79HF6 cells and LAN-1 cells were also irradiated by Carbon-UNILAC with LET of 168?keV/μm and Nickel ions with LET of 174?keV/μm, respectively. The effect of radiation on the survival and proliferation of cells was addressed by standard clonogenic assays. In order to analyze cell karyotype standard Giemsa staining, multicolor fluorescence in situ hybridization (mFISH) and multicolor banding (mBAND) techniques were applied. Results Relative biological effectiveness values of heavy ion beams relative to X-rays at the D_(10)values were found between 2.3 and 2.6 with Carbon-SIS and Nickel for LAN-1 and between 2.5 and 3.4 with Carbon-SIS and Carbon-UNILAC for 79HF6 cells. Chemorefractory LAN-1~(RETO)cells were found more radioresistant than untreated LAN-1~(WT)cells. 79HF6~(RETO)glioblastoma cells were found more radiosensitive than cytostatic sensitive cells 79HF6~(WT). Sphere formation assay showed that LAN-1~(RETO)cells were able to form spheres in serum-free culture, whereas 79HF6 cells could not. Most of 79HF6~(WT)cells revealed a number of 71–90 chromosomes, whereas 79HF6~(RETO)revealed a number of 52–83 chromosomes. The majority of LAN-1~(WT)cells revealed a number of 40–44 chromosomes. mFISH analysis showed some stable aberrations, especially on chromosome 10 as judged by the impossibility to label this region with specific probes. This was corroborated using mBAND analysis. Conclusion Heavy ion irradiation was more effective than X-ray in both cytostatic naive cancer and chemoresistant cell lines. LAN-1~(RETO)chemoresistant neuroblastoma cells were found to be more radioresistant than the cytostatic naive cells (LAN-1~(WT)), whereas this effect was not found in 79HF6 cells.
机译：目的这项研究的目的是将数字和结构染色体畸变与放疗在化疗难治性肿瘤细胞中的有效性联系起来。材料和方法将来自患者及其化学抗性亚系的神经母细胞瘤（LAN-1）和79HF6胶质母细胞瘤细胞作为神经球人工培养，并用X射线和重离子源照射。所有细胞系均用Carbon-SIS辐照，LET为100？keV /μm。但是，Carbon-UNILAC还分别用LET 168？keV /μm和LET 174？keV /μm的镍离子辐照了79HF6细胞和LAN-1细胞。辐射对细胞存活和增殖的影响已通过标准克隆形成试验得以解决。为了分析细胞核型标准Giemsa染色，应用了多色荧光原位杂交（mFISH）和多色条带（mBAND）技术。结果对于Carbon-SIS和镍，对于LAN-1，重离子束相对于X射线在D_（10）值的相对生物有效性值在2.3和2.6之间;对于Carbon-SIS和Carbon-UNILAC，在2.5和3.4之间适用于79HF6细胞。发现化学难治性LAN-1〜（RETO）细胞比未处理的LAN-1〜（WT）细胞更耐辐射。发现79HF6〜（RETO）胶质母细胞瘤细胞比细胞抑制敏感性细胞79HF6〜（WT）更具放射敏感性。球体形成实验表明，LAN-1〜（RETO）细胞能够在无血清培养物中形成球体，而79HF6细胞则不能。大多数79HF6〜（WT）细胞揭示了71-90条染色体，而79HF6〜（RETO）揭示了52-83条染色体。大多数LAN-1〜（WT）细胞显示40-44条染色体。 mFISH分析显示出一些稳定的像差，尤其是在第10号染色体上，这是由于无法用特定探针标记该区域而判断的。使用mBAND分析证实了这一点。结论重离子辐射对原发性癌细胞和化疗耐药细胞系均优于X射线。发现LAN-1〜（RETO）化学抗性神经母细胞瘤细胞比具有细胞生长抑制作用的幼稚细胞（LAN-1〜（WT））具有更高的放射抗性，而在79HF6细胞中则没有这种作用。

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