Brady-Kalnay S M, Boghaert E R, Zimmer S, Brackenbury R
Department of Anatomy and Cell Biology, University of Cincinnati Medical Center, OH 45267-0521.
Clin Exp Metastasis. 1993 Jul;11(4):313-24. doi: 10.1007/BF00058051.
The WC5 rat cerebellar cell line, infected with a Rous sarcoma virus (RSV) that is temperature-sensitive for pp60v-src transformation, expresses high levels of the neural cell adhesion molecule, N-CAM, when grown at the non-permissive temperature for pp60v-src activity. At the permissive temperature, N-CAM expression is 4- to 10-fold reduced and the cells aggregate poorly. To evaluate the effects of variations in N-CAM expression, we compared the invasive ability of transformed WC5 cells that express low levels of N-CAM with transformed cells in which N-CAM-mediated adhesion was restored. WC5 cells were transfected with expression vectors containing cDNAs encoding the 120 or 180 kDa forms of chicken N-CAM linked to constitutive promoters. Several permanently transfected lines that expressed chicken N-CAM at the cell surface were isolated. These cell lines showed enhanced aggregation at the permissive temperature relative to untransfected WC5 cells or cells transfected with control constructs. By comparing the ability of control and transfected WC5 cells to invade reconstituted extracellular matrix, we tested the effect of variations in N-CAM-mediated adhesion on invasion. Clones that expressed high levels of N-CAM showed invasion rates that were similar to control cells, indicating that increasing N-CAM-mediated adhesion does not inhibit the invasiveness of RSV-transformed WC5 cells.
WC5大鼠小脑细胞系感染了对pp60v-src转化具有温度敏感性的劳氏肉瘤病毒(RSV),当在pp60v-src活性的非允许温度下生长时,该细胞系表达高水平的神经细胞粘附分子N-CAM。在允许温度下,N-CAM表达降低4至10倍,细胞聚集性差。为了评估N-CAM表达变化的影响,我们比较了表达低水平N-CAM的转化WC5细胞与恢复了N-CAM介导粘附的转化细胞的侵袭能力。用含有与组成型启动子相连的编码120或180 kDa形式鸡N-CAM的cDNA的表达载体转染WC5细胞。分离出几个在细胞表面表达鸡N-CAM的永久转染细胞系。相对于未转染的WC5细胞或用对照构建体转染的细胞,这些细胞系在允许温度下显示出增强的聚集性。通过比较对照和转染的WC5细胞侵入重组细胞外基质的能力,我们测试了N-CAM介导的粘附变化对侵袭的影响。表达高水平N-CAM的克隆显示出与对照细胞相似的侵袭率,表明增加N-CAM介导的粘附并不抑制RSV转化的WC5细胞的侵袭性。