Department of Neurological Surgery, University of California at San Francisco, San Francisco, California 94143, USA.
J Clin Neurosci. 2010 Dec;17(12):1573-7. doi: 10.1016/j.jocn.2010.05.007. Epub 2010 Sep 9.
Alteration of tumor cell growth kinetics is the goal of nearly all current or proposed therapies for human neoplasms. The adenosine triphosphate (ATP) chemiluminescent assay has been used for some time as a surrogate marker of in vitro cell growth. Here we present data showing that three human glioblastoma cell lines (U87, U251, G55) demonstrate significantly different cell number to luminescence relationships when subjected to this assay. We plated progressively increasing numbers of cells per well; from 1000 to 50,000 were grown in Dulbecco's modified Eagle's medium without serum and cultured for 6 hours. Cells were then lysed and subjected to the chemiluminescent assay to measure ATP levels and a linear relationship between cell number and measured luminescence was found. Despite this, we found that the slope of the regression line (β) varied markedly between different cell lines (U251 [β = 0.968 ± 0.3] vs. U87 [β = 0.772 ± 0.2] vs. G55 [β = 0.757 ± 0.2]; p < 0.0001), suggesting a difference in ATP luminescence per cell between these cell lines. Thus, we have demonstrated that luminescence values are internally linear within a given cell population, but luminescence level per cell varies significantly between different glioma cell lines. Our findings suggest that different glioma cell lines have unique levels of ATP per cell.
改变肿瘤细胞生长动力学是目前几乎所有人类肿瘤治疗方法的目标。三磷酸腺苷（ATP）化学发光测定法已被用于一段时间，作为体外细胞生长的替代标志物。这里我们提供的数据显示，三种人神经胶质瘤细胞系（U87、U251、G55）在接受这种测定时表现出明显不同的细胞数与发光之间的关系。我们在每个孔中逐渐增加细胞数量；从 1000 到 50000 在无血清的 Dulbecco 修改的 Eagle 培养基中生长，并培养 6 小时。然后将细胞裂解并进行化学发光测定以测量 ATP 水平，并发现细胞数与测量的发光之间存在线性关系。尽管如此，我们发现不同细胞系之间回归线（β）斜率差异显著（U251 [β = 0.968 ± 0.3] 与 U87 [β = 0.772 ± 0.2] 与 G55 [β = 0.757 ± 0.2]；p < 0.0001），表明这些细胞系之间每个细胞的 ATP 发光差异。因此，我们已经证明在给定的细胞群体内发光值是内部线性的，但是每个细胞的发光水平在不同的神经胶质瘤细胞系之间有很大的差异。我们的研究结果表明，不同的神经胶质瘤细胞系具有独特的每个细胞的 ATP 水平。
