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首页> 外文期刊>Radiotherapy and oncology: Journal of the European Society for Therapeutic Radiology and Oncology >Selection of radioresistant tumor cells and presence of ALDH1 activity in vitro.
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Selection of radioresistant tumor cells and presence of ALDH1 activity in vitro.

机译：体外放射抗肿瘤细胞的选择和ALDH1活性的存在。

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摘要

BACKGROUND: Tumor resistance to radiotherapy has been hypothesized to be mediated by a tumor subpopulation, called cancer stem cells (CSCs). Based on the proposed function of CSCs in radioresistance, we explored the cancer stem cell properties of cells selected for radioresistance phenotype. MATERIALS AND METHODS: A549 and SK-BR-3 cells were radioselected with four single doses of 4 or 3 Gy in intervals of 10-12 days and used for colony formation assay and gamma-H2AX foci formation assay. Expression of putative stem cell markers, i.e. Sox2, Oct4, ALDH1, and CD133 were analyzed using Western blotting. A549 and SK-BR-3 cells sorted based on their ALDH1 activity were analyzed in clonogenic survival assays. RESULTS: Radioselected A549 and SK-BR-3 cells (A549-R, SK-BR-3-R) showed increased radioresistance and A549-R cells presented enhanced repair of DNA-double strand breaks. PI3K inhibition significantly reduced radioresistance of A549-R cells. Cell line specific differences in the expression of the putative CSC markers Sox2 and Oct4 were observed when parental and radioselected cells were compared but could not be directly correlated to the radioresistant phenotype. However, enzyme activity of the putative stem cell marker ALDH1 showed a correlation to radioresistance. CONCLUSIONS: Subpopulations of pooled radioresistant colonies, selected by various radiation exposures were analyzed for the presence of putative stem cell markers. Although the pattern of Sox2, Oct4, and CD133 expression was not generally associated with radioresistance, presence of ALDH1 seems to be indicative for subpopulations with increased radioresistance.
机译：背景：已经假设肿瘤对放射疗法的抗性是由称为癌症干细胞（CSC）的肿瘤亚群介导的。基于拟议中的CSCs在放射抗性中的功能，我们探索了选择用于放射抗性表型的细胞的癌症干细胞特性。材料与方法：在10-12天的间隔内，以4或3 Gy的四次单剂量对A549和SK-BR-3细胞进行放射选择，并用于集落形成测定和γ-H2AX灶形成测定。使用蛋白质印迹分析推定的干细胞标志物，即Sox2，Oct4，ALDH1和CD133的表达。在克隆形成存活分析中分析了根据其ALDH1活性分类的A549和SK-BR-3细胞。结果：放射选择的A549和SK-BR-3细胞（A549-R，SK-BR-3-R）显示出增加的抗辐射性，而A549-R细胞表现出增强的DNA双链断裂修复。 PI3K抑制显着降低了A549-R细胞的放射抗性。当比较亲代和放射选择的细胞时，观察到推定的CSC标记Sox2和Oct4表达的细胞系特异性差异，但不能与放射抗性表型直接相关。但是，推定的干细胞标志物ALDH1的酶活性与抗辐射性相关。结论：通过各种辐射暴露选择了合并的放射抗性菌落的亚群，分析了推定的干细胞标记物的存在。尽管Sox2，Oct4和CD133的表达模式通常与抗辐射性无关，但ALDH1的存在似乎表明抗辐射性增强的亚群。

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