用流式细胞术检测γ射线诱发淋巴细胞γ-H2AX与照射剂量的关系

Relationship between γ-H2AX in lymphocytes and radiation dose of γ-rays by flow cytometry assay

本研究使用流式细胞术检测了外周血淋巴细胞γ-H2AX表达水平,探讨了该方法用于辐射生物剂量估算的可能性。采集健康人外周血,进行体外60Coγ射线照射,利用γ-H2AX特异性抗体对淋巴细胞进行间接免疫荧光标记,采用流式细胞仪分析淋巴细胞中γ-H2AX的几何平均荧光强度值,并计算出相对荧光强度;比较了仪器类型和参数设定对上述两项指标的影响,以上述两项指标分别建立照射后30 min的剂量-效应曲线并进行对比。结果显示:参数设定和仪器类型均明显影响60Coγ射线照射后γ-H2AX几何平均荧光强度,而对相对荧光强度无显著影响。因此,用流式细胞仪分析γ-H2AX的相对荧光强度,较好地解决了实验的稳定性和重复性问题,有望成为一种估算辐射生物剂量的新方法。

To search for the opportunity for assessing doses as a biological dosimeter, an approach was estab- lished for detecting the expression levels of γ-H2AX in peripheral blood lymphocytes by flow cytometry. The periph- eral blood from health donors irradiated in vitro by 60Co y-rays. The geometric mean fluorescence intensity values of y-H2AX in lymphocytes labeled with anti-γ-H2AX-antibody by using indirect immunofluorescence were analyzed by flow cytometry, and the relative fluorescence intensity was calculated. The geometric mean fluorescence intensity values and the relative fluorescence intensity of γ-H2AX in intracellular effect were compared between the different instruments type and parameters setting. Two dose-response curves were established and compared 30 rain post-irradiated when the geometric mean fluorescence intensity and the relative fluorescence intensity of γ-H2AX in lymphocytes were used as the indicators. The geometric mean fluoreseenee intensity was obviously affected by different instruments type and parameters setting after irradiation with various doses, but the relative fluorescence intensity of γ-H2AX in lymphocytes did not significantly correlate with the different instruments type and parameters set-ting after irradiation with various doses. The relative fluorescence intensity as the indicator analyzed by flow cytometry might improve the stability and repeatability, so this new approach would be a potential biological dosimeter.