Ben-Ze'ev A, Reiss R, Bendori R, Gorodecki B
Department of Molecular Genetics and Virology, Weizmann Institute of Science, Rehovot, Israel.
Cell Regul. 1990 Aug;1(9):621-36. doi: 10.1091/mbc.1.9.621.
When stimulated with serum, quiescent Balb/C-3T3 fibroblasts were found to induce vinculin transcription transiently within 30 min, followed by accumulation of vinculin mRNA and protein synthesis between 2 and 4 h after stimulation and a decrease to the basal level by 6-8 h. Platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), and 12-O-tetradecanoylphorbol-13-acetate (TPA) each could elicit a similar response, albeit to a lesser extent, whereas epidermal growth factor (EGF) was inefficient in inducing vinculin expression. In cells stimulated with serum and cycloheximide, vinculin expression was superinduced and vinculin mRNA levels persisted longer than in cells stimulated with serum alone. Cells arrested in the presence of serum by anchorage denial in methyl cellulose suspension culture also induced vinculin expression and formed large vinculin positive plaques when reattaching and spreading on the substrate in the presence of serum. Cells replated from suspension culture in the absence of serum on either plastic or extracellular matrix (ECM) components were capable of extensive spreading, but failed to elevate vinculin expression and displayed diffuse vinculin staining. The results indicate that the changes in vinculin organization and expression in response to growth factor stimulation may reflect either a necessary step in the progression through the cell cycle or a response related to complex cellular processes such as wound repair and embryogenesis.
当用血清刺激时,发现静止的Balb/C - 3T3成纤维细胞在30分钟内短暂诱导纽蛋白转录,随后在刺激后2至4小时内纽蛋白mRNA积累和蛋白质合成,到6 - 8小时降至基础水平。血小板衍生生长因子(PDGF)、成纤维细胞生长因子(FGF)和12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯(TPA)各自都能引发类似反应,尽管程度较小,而表皮生长因子(EGF)诱导纽蛋白表达的效率较低。在用血清和环己酰亚胺刺激的细胞中,纽蛋白表达被超诱导,且纽蛋白mRNA水平持续的时间比仅用血清刺激的细胞更长。通过在甲基纤维素悬浮培养中剥夺贴壁条件而在血清存在下停滞的细胞,在血清存在下重新附着并铺展在底物上时也诱导纽蛋白表达并形成大的纽蛋白阳性斑块。在无血清条件下从悬浮培养物重新接种到塑料或细胞外基质(ECM)成分上的细胞能够广泛铺展,但未能提高纽蛋白表达并显示出弥漫性的纽蛋白染色。结果表明,响应生长因子刺激时纽蛋白组织和表达的变化可能反映了细胞周期进程中的一个必要步骤,或者是与伤口修复和胚胎发生等复杂细胞过程相关的一种反应。