Skarsgard L D, Wilson D J, Durand R E
Department of Medical Biophysics, B.C. Cancer Research Centre, Vancouver, Canada.
Radiat Res. 1993 Jan;133(1):102-7.
In an earlier study using cell sorting techniques to define the radiation survival response of asynchronous Chinese hamster V79-171 cells more accurately, we found evidence of substructure in the response at low dose. In the present work we have attempted to show that this substructure arises from the subpopulations of sensitive (G1, G2 phase) and resistant (late S phase) cells which are present in asynchronously dividing cultures but which are not resolved by conventional survival assays. Partially synchronized cells were produced by exposure to 1 mM hydroxyurea for 12 h and were harvested 15 min later, yielding a population of viable cells at or just beyond the G1/S-phase boundary. Parallel experiments were carried out with asynchronous cells. The average of repeated measurements of the radiation survival response of asynchronous cells again showed a significant difference (P = 0.002 to 0.009) between the alpha and beta values evaluated from the data for the low-dose range, 0-2.8 Gy, and the high-dose range, 2.8-14 Gy. For G1/S-phase cells, however, there was no significant difference between the values of alpha and beta for the low-dose and high-dose ranges (P > 0.5). The results thus support the hypothesis that the observed substructure in the asynchronous response is due to resolution of subpopulations of different radiosensitivities, and they illustrate the advantage of the cell sorter assay for accurate measurements of cell survival, particularly at low dose.
在一项早期研究中,我们使用细胞分选技术更准确地定义异步培养的中国仓鼠V79 - 171细胞的辐射存活反应,发现在低剂量反应中存在亚结构的证据。在本研究中,我们试图表明这种亚结构源于敏感细胞(G1期、G2期)和抗性细胞(S期后期)的亚群,这些亚群存在于异步分裂培养物中,但传统的存活分析无法分辨。通过暴露于1 mM羟基脲12小时产生部分同步化细胞,并在15分钟后收获,得到处于或刚超过G1/S期边界的活细胞群体。对异步细胞进行了平行实验。对异步细胞辐射存活反应的重复测量平均值再次显示,从低剂量范围(0 - 2.8 Gy)和高剂量范围(2.8 - 14 Gy)的数据评估的α和β值之间存在显著差异(P = 0.002至0.009)。然而,对于G1/S期细胞,低剂量和高剂量范围的α和β值之间没有显著差异(P > 0.5)。因此,结果支持了这样的假设,即异步反应中观察到的亚结构是由于不同放射敏感性亚群的分辨,并且它们说明了细胞分选仪测定法在准确测量细胞存活方面的优势,特别是在低剂量时。