St-Arnaud R, Craig J, McBurney M W, Papkoff J
Department of Medicine, University of Ottawa, Canada.
Oncogene. 1989 Sep;4(9):1077-80.
We have used the P19 line of mouse embryonal carcinoma (EC) cells to initiate studies on the putative role of the int-1 proto-oncogene during neuronal differentiation. P19 cells are induced to differentiate into neurons, astrocytes and fibroblast-like cells following exposure to retinoic acid (RA). Treatment of the same P19 cells with dimethyl sulfoxide (DMSO) leads to differentiation into mesodermal derivatives, including skeletal and cardiac muscle. Northern blot analysis showed that int-1 RNA was not present in the EC cells but appeared 48 h after RA exposure and could be detected for at least the next 8 days. No int-1 RNA was detected in P19 cells induced to differentiate with DMSO. Nuclear run-on transcription assays showed that int-1 expression in RA-treated P19 cells was induced at the transcriptional level. Immunofluorescent staining with an antibody directed against an int-1 peptide identified immunoreactive material in cytoplasmic granules of fibroblast-like cells in RA-treated P19 cultures. Thus the P19 cell line is a suitable experimental system to study int-1 gene expression and function during neuroectodermal development.
我们利用小鼠胚胎癌细胞系P19来开展关于int-1原癌基因在神经元分化过程中假定作用的研究。将P19细胞暴露于视黄酸（RA）后，可诱导其分化为神经元、星形胶质细胞和成纤维细胞样细胞。用二甲基亚砜（DMSO）处理同样的P19细胞，则会使其分化为中胚层衍生物，包括骨骼肌和心肌。Northern印迹分析表明，int-1 RNA在胚胎癌细胞中并不存在，但在暴露于RA后48小时出现，并且在接下来至少8天内都能检测到。在用DMSO诱导分化的P19细胞中未检测到int-1 RNA。细胞核连续转录分析表明，经RA处理的P19细胞中int-1的表达是在转录水平上被诱导的。用针对int-1肽段的抗体进行免疫荧光染色，在经RA处理的P19培养物中成纤维细胞样细胞的细胞质颗粒中鉴定出了免疫反应性物质。因此，P19细胞系是研究神经外胚层发育过程中int-1基因表达和功能的合适实验系统。
