Procaspase 3 activator compounds combined with X-ray irradiation

我们已经从我们在伊利诺伊大学的合作者那里获得了两个proaspase 3激活剂(PAC-1和SPAC-1)，并获得了一组细胞株，在这些细胞系上测试化合物在X射线照射和不照射的情况下。对一些肉瘤细胞株的初步实验表明，当以低密度培养时，它们不会形成离散的集落，这表明评估辐射效应的金标准(集落形成实验)不能直接应用。我们已经评估了其他几种基于磷酸化的伽马-H_2AX组蛋白的分析方法，以量化在没有和存在原天冬氨酸酶3激活剂的情况下辐射的影响。此外，我们正在验证使用基于ATP的检测来确定细胞系中的放射增敏活性。这已经取得了显著的进展，我们已经开始对细胞系进行最终的分析。这揭示了几个有希望的结果。我们还在同时测试正在研究的细胞系中的凋亡机制的功能。我们目前正在验证每个肉瘤细胞系的这些检测方法。

We have obtained two procaspase 3 activators (PAC-1 and sPAC-1) from our collaborators at the University of Illinois, and have obtained a panel of cell lines on which to test the compounds with and without X-irradiation. Initial experiments with a number of sarcoma cell lines showed that they did not form discrete colonies when plated at low density, indicating that the gold standard evaluation of the effects of radiation (colony formation assay) could not be applied directly. We have evaluated several other assays based on phosphorylated gamma-H2AX histone proteins to quantitate the effect of radiation in both the absence and presence of procaspase 3 activators. In addition, we are validating the use of an ATP-based assay to determine radiosensitizing activity in cell lines. This has progressed significantly, and we have begun definitive assays on the cell lines. This has revealed several promising results. We are also concurrently testing the function of apoptotic machinery in cell lines under investigation. We are currently validating these assays for each sarcoma cell line.