Chang Yu-Ying, Chen Shiang-Jiuun, Liang Huang-Chien, Sung Hsing-Wen, Lin Chien-Chung, Huang Rong-Nan
Department of Life Science, National Central University, Chung-Li, 32054 Taoyuan, Taiwan, Republic of China.
Biomaterials. 2004 Aug;25(17):3603-11. doi: 10.1016/j.biomaterials.2003.10.039.
Galectin-1 (GAL1), a beta-galactoside-binding protein, functions in cell adhesion, development, and growth regulation. A number of studies suggest that GAL1 play an important role in enhancing cell adhesion to extracellular matrix and inducing cell proliferation. Chitosan is a derivative of chitin extracted from lobsters, crabs and shrimps' exoskeletons. In clinical medicine, chitosan membrane had been used as a semi-permeable biological dressing. Although chitosan membranes show no cytotoxicity, some cell types (e.g. 3T3 cells) fail to attach and proliferate on their surface. In these studies, we show that over-expression of GAL1 does not enhance 3T3 cell proliferation on chitosan membranes. However, coating the chitosan membrane with recombinant GAL1 proteins significantly expedites 3T3 cells proliferation. The enhanced cell growth was inhibited by thiodigalactoside (TDG, a potent inhibitor of beta-galactoside binding) and GAL1 monoclonal antibodies, suggesting GAL1's specific effect on the proliferation of 3T3 cells upon chitosan membranes. Moreover, immunoblotting detected a markedly suppressed tyrosine phosphorylation in several proteins on 3T3 cell growths upon GAL1-coated chitosan membrane. Pretreating the cells with sodium fluoride (NaF, a phosphatase inhibitor) inhibits the attachment and proliferation of 3T3 cells. These findings support a proposed role for altered levels of protein phosphorylation in GAL1-mediated cell attachment and proliferation on chitosan membranes.
半乳糖凝集素-1(GAL1)是一种β-半乳糖苷结合蛋白,在细胞黏附、发育和生长调节中发挥作用。多项研究表明,GAL1在增强细胞与细胞外基质的黏附以及诱导细胞增殖方面发挥重要作用。壳聚糖是从龙虾、螃蟹和虾的外骨骼中提取的几丁质的衍生物。在临床医学中,壳聚糖膜已被用作半透性生物敷料。尽管壳聚糖膜没有细胞毒性,但某些细胞类型(如3T3细胞)无法在其表面附着和增殖。在这些研究中,我们表明GAL1的过表达并不能增强3T3细胞在壳聚糖膜上的增殖。然而,用重组GAL1蛋白包被壳聚糖膜可显著加速3T3细胞的增殖。硫代二半乳糖苷(TDG,一种有效的β-半乳糖苷结合抑制剂)和GAL1单克隆抗体可抑制细胞生长的增强,这表明GAL1对壳聚糖膜上3T3细胞的增殖具有特异性作用。此外,免疫印迹检测到在GAL1包被的壳聚糖膜上3T3细胞生长时几种蛋白质中的酪氨酸磷酸化明显受到抑制。用氟化钠(NaF,一种磷酸酶抑制剂)预处理细胞可抑制3T3细胞的附着和增殖。这些发现支持了蛋白质磷酸化水平改变在GAL1介导的壳聚糖膜上细胞附着和增殖中所起作用的观点。