Bao Xingfeng, Fukuda Minoru
Sanford-Burnham Medical Research Institute, La Jolla, California, USA.
Methods Enzymol. 2010;479:387-96. doi: 10.1016/S0076-6879(10)79022-4.
Interaction of epithelial cells with basement membrane (BM) is mediated by cell-adhesion molecules, which regulate cell proliferation, motility, and differentiation by integrating signals from extracellular matrix and soluble factors. alpha-Dystroglycan (alpha-DG) is one of the most important adhesion molecules in epithelial cell-BM interaction. alpha-DG serves as the cell surface receptor for several major BM proteins, including laminin, perlecan, and agrin. The laminin G-like domain in all these proteins binds to a unique glycan structure, so-called laminin-binding glycan, attached to alpha-DG with high affinity. Formation of the laminin-binding glycan is required for the BM assembly, and loss or deficiency of the glycan causes muscular dystrophy. We studied the role of this alpha-DG-specific glycan modification in tumor development, and identified a tumor suppressor function of the laminin-binding alpha-DG. In this chapter, we describe methods used to isolate the cell populations from human prostate cancer cell line PC3 and characterize their potentials in tumor formation and metastasis in vitro and in vivo.
上皮细胞与基底膜（BM）的相互作用由细胞粘附分子介导，这些分子通过整合来自细胞外基质和可溶性因子的信号来调节细胞增殖、运动和分化。α- dystroglycan（α-DG）是上皮细胞与基底膜相互作用中最重要的粘附分子之一。α-DG作为几种主要基底膜蛋白的细胞表面受体，包括层粘连蛋白、基底膜聚糖和聚集蛋白。所有这些蛋白中的层粘连蛋白G样结构域与一种独特的聚糖结构（即所谓的层粘连蛋白结合聚糖）以高亲和力结合，该聚糖附着于α-DG。层粘连蛋白结合聚糖的形成是基底膜组装所必需的，聚糖的缺失或缺陷会导致肌肉萎缩症。我们研究了这种α-DG特异性聚糖修饰在肿瘤发生中的作用，并确定了层粘连蛋白结合α-DG的肿瘤抑制功能。在本章中，我们描述了从人前列腺癌细胞系PC3中分离细胞群体的方法，以及在体外和体内表征它们在肿瘤形成和转移方面的潜力的方法。
