Trotter J, Boulter C A, Sontheimer H, Schachner M, Wagner E F
Department of Neurobiology, University of Heidelberg, Federal Republic of Germany.
Oncogene. 1989 Apr;4(4):457-64.
A replication-defective retroviral vector carrying the v-src oncogene and the gene for neomycin resistance was used to infect neurone- and fibroblast-depleted embryonic mouse brain cells in vitro. Cells resistant to the antibiotic G418 were obtained and continually passaged. Several cell lines were isolated which express high levels of v-src mRNA and v-src tyrosine kinase activity. The antigenic marker profile of either the pooled cells from an individual infection or sublines isolated from individual foci showed the cells to be immature glia: most cells expressed vimentin, A2B5 antigen and/or J1/tenascin glycoproteins, but not fibronectin. Sublines expressed different antigen profiles suggesting that the immortalised cells were derived from glial cells of different phenotypes. The cell lines expressed the 120 and 140 but not the 180 kd components of N-CAM as well as voltage-activated potassium channels, typical for glial cells. 01 antigen-positive oligodendrocytes were never observed in the lines or sublines after long term passage (over 1 year), but some cells expressed glial fibrillary acidic protein, a marker for mature astrocytes. Thus, expression of v-src in murine glial cells appears to arrest their development and prevent their differentiation.
一种携带v-src癌基因和新霉素抗性基因的复制缺陷型逆转录病毒载体，被用于在体外感染去除了神经元和成纤维细胞的胚胎小鼠脑细胞。获得了对抗生素G418有抗性的细胞，并持续传代培养。分离出了几个细胞系，它们表达高水平的v-src mRNA和v-src酪氨酸激酶活性。来自单个感染的混合细胞或从单个病灶分离出的亚系的抗原标志物谱显示，这些细胞是未成熟的神经胶质细胞：大多数细胞表达波形蛋白、A2B5抗原和/或J1/腱生蛋白糖蛋白，但不表达纤连蛋白。亚系表达不同的抗原谱，表明永生化细胞来源于不同表型的神经胶质细胞。这些细胞系表达N-CAM的120和140 kd成分，但不表达180 kd成分，以及电压激活钾通道，这是神经胶质细胞的典型特征。长期传代（超过1年）后，在这些细胞系或亚系中从未观察到01抗原阳性的少突胶质细胞，但一些细胞表达胶质纤维酸性蛋白，这是成熟星形胶质细胞的标志物。因此，v-src在小鼠神经胶质细胞中的表达似乎会阻止它们的发育并防止其分化。
