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目的硼中子俘获疗法(BNCT)的有效性主要取决于硼-10元素在肿瘤细胞内的聚集。目前的硼检测手段仅限于分析肿瘤组织中硼的平均浓度,不能准确地评价硼化合物(BPA)在组织中的详细分布。先前研究证实温热处理可以提高放疗疗效,本研究通过免疫荧光法检测温热是否对BPA细胞内的微分布有影响。方法通过免疫荧光法分析硼化合物在肿瘤细胞中的分布以及温热对BPA分布的影响。在体外对人肺癌细胞株A549和人头颈部鳞状细胞癌细胞株SAS进行培养,并且加入BPA共培养1h。另外,A549细胞经温热处理1h后,利用抗BPA抗体通过免疫荧光法进行检测。免疫荧光强度通过图像分析软件进行分析。结果检测发现BPA主要聚集于细胞核周围,BPA免疫荧光的强度随着培养基中BPA浓度的增加而增强。尽管在26μg/ml浓度和13μg/ml浓度时,细胞免疫荧光的平均强度相近,但直方图分析发现26μg/ml时细胞主要向高荧光强度区域移动。经温热处理后,BPA在细胞中的分布没有明显增加。结论本研究利用免疫荧光法对BPA在细胞内的分布进行了分析,以上结果暗示温热对肿瘤细胞硼摄取的影响可能主要需要通过改善肿瘤循环、增加肿瘤内BPA的输送来实现。
Purpose The effectiveness of boron neutron capture therapy (BNCT) depends primarily on the aggregation of boron-10 elements within tumor cells. Current boron detection methods are limited to the analysis of the average concentration of boron in tumor tissue and do not accurately assess the detailed distribution of boron compounds (BPA) in tissues. Previous studies have confirmed that warm treatment can improve the efficacy of radiotherapy, the study by immunofluorescence test whether the warmth of BPA intracellular micro distribution affect. Methods The distribution of boron compounds in tumor cells and the effect of warming on BPA distribution were analyzed by immunofluorescence. Human lung cancer cell line A549 and human head and neck squamous cell carcinoma cell line SAS were cultured in vitro and co-cultured with BPA for 1 hour. In addition, after A549 cells were treated with warm treatment for 1 h, their anti-BPA antibodies were detected by immunofluorescence. Immunofluorescence intensity was analyzed by image analysis software. The results showed that BPA mainly gathered around the nucleus and the intensity of BPA immunofluorescence increased with the increase of BPA concentration in the medium. Although the average intensity of cellular immunofluorescence was similar at 26 and 13 μg / ml, the histogram analysis found that the cells mainly moved to the high fluorescence intensity region at 26 μg / ml. After warming, the distribution of BPA in the cells did not increase significantly. Conclusions In this study, immunofluorescence assay was used to analyze the intracellular distribution of BPA. The above results suggest that the influence of warmth on the uptake of boron by tumor cells may be mainly achieved by improving tumor circulation and increasing intra-tumor BPA delivery.