Clark G J, Cox A D, Graham S M, Der C J
Department of Pharmacology, School of Medicine, University of North Carolina at Chapel Hill 27599, USA.
Methods Enzymol. 1995;255:395-412. doi: 10.1016/s0076-6879(95)55042-9.
The rodent fibroblast systems described above have provided sensitive and rapid biological assays to characterize the properties of normal and mutated Ras proteins. Furthermore, these assays have provided in vitro systems to measure the ability of other cellular components to modulate Ras signal transduction and transformation. However, while these assays provide an excellent measure of Ras-transforming activity, the fact that these cells are of fibroblastic origin, and can be transformed by a single hit, indicates that caution should be used in extrapolating observations from NIH 3T3 transformation assays to the situation in human tumors. Therefore, using human epithelial cell-based assays that more closely approximate the cell types where mutated ras alleles are most frequently detected may provide more realistic assays for examining the biochemical and biological consequences of aberrant Ras function in human tumors. Nevertheless, despite these cautions, these rodent transformation assays will continue to be the best and most widely applied assays for Ras biological activity.
上述啮齿动物成纤维细胞系统提供了灵敏且快速的生物学检测方法，以表征正常和突变型Ras蛋白的特性。此外，这些检测方法还提供了体外系统，用于测量其他细胞成分调节Ras信号转导和转化的能力。然而，尽管这些检测方法能很好地衡量Ras转化活性，但这些细胞起源于成纤维细胞，且只需一次打击就能被转化，这一事实表明，在将NIH 3T3转化检测的观察结果外推至人类肿瘤情况时应谨慎。因此，使用基于人类上皮细胞的检测方法，这些方法更接近最常检测到突变型ras等位基因的细胞类型，可能会为研究异常Ras功能在人类肿瘤中的生化和生物学后果提供更现实的检测方法。尽管如此，尽管有这些注意事项，这些啮齿动物转化检测仍将继续是用于Ras生物学活性的最佳且应用最广泛的检测方法。
