Hatai M, Hashi H, Kato I, Yaoi Y
Biology Division, National Cancer Center Research Institute, Tokyo, Japan.
Cell Struct Funct. 1993 Feb;18(1):53-60. doi: 10.1247/csf.18.53.
Type V collagen inhibits the cell-substratum adhesion of many types of cells. In this study, inhibitory effects of type V collagen on the adhesion of mouse melanoma B16-F10 cells to fibronectin, laminin and vitronectin were investigated. When the culture dishes were coated with a mixture of fibronectin and type V collagen, adhesion of the cells was inhibited by 50% at a fibronectin/collagen molar ratio of 10/1. At a similar molar ratio, adhesion of the cells to laminin was inhibited moderately, but that to vitronectin was not significantly affected. Type V collagen added into culture medium was less effective in inhibiting cell adhesion. The antiadhesive activity of type V collagen was partially retained in the alpha 1 (V) chain of heat-denatured collagen. The alpha 1 (V) chain was split into two large fragments, 90 kDa and 60 kDa, by limited digestion with Staphylococcus aureus V8 proteinase. The 90-kDa fragment, which was derived from the C-terminal half of the alpha 1 (V) chain, inhibited the cell adhesion more profoundly than alpha 1 (V). However, little fibronectin bound to the 90-kDa fragment, while fibronectin bound to the 60-kDa fragment, which was less antiadhesive than the 90-kDa fragment, with the same extent as alpha 1 (V). We therefore concluded that the antiadhesive effect of type V collagen was not due to its specific binding to the fibronectin molecule.
V型胶原可抑制多种细胞与细胞外基质的黏附。在本研究中，我们调查了V型胶原对小鼠黑色素瘤B16-F10细胞与纤连蛋白、层粘连蛋白和玻连蛋白黏附的抑制作用。当培养皿用纤连蛋白和V型胶原的混合物包被时，在纤连蛋白/胶原摩尔比为10/1时，细胞黏附被抑制了50%。在相似的摩尔比下，细胞与层粘连蛋白的黏附受到适度抑制，但与玻连蛋白的黏附没有受到显著影响。添加到培养基中的V型胶原在抑制细胞黏附方面效果较差。热变性胶原的α1(V)链部分保留了V型胶原的抗黏附活性。通过金黄色葡萄球菌V8蛋白酶的有限酶切，α1(V)链被切割成两个大片段，90 kDa和60 kDa。源自α1(V)链C端一半的90 kDa片段比α1(V)更能深刻地抑制细胞黏附。然而，几乎没有纤连蛋白与90 kDa片段结合，而纤连蛋白与60 kDa片段结合，60 kDa片段的抗黏附性比90 kDa片段弱，其与纤连蛋白的结合程度与α1(V)相同。因此，我们得出结论，V型胶原的抗黏附作用不是由于其与纤连蛋白分子的特异性结合。
