Yamada K M, Kennedy D W, Yamada S S, Gralnick H, Chen W T, Akiyama S K
Membrane Biochemistry Section, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Cancer Res. 1990 Aug 1;50(15):4485-96.
The processes of migration and invasion by human tumor cells are likely to involve specific cell surface receptors, such as receptors for the extracellular matrix molecules fibronectin, laminin, and collagen. We have examined the roles of several of these receptors using a set of monoclonal antibodies directed against the beta 1 integrin family, as well as a series of synthetic peptides reported to inhibit various interactions of each of these proteins with the cell surface. The most general inhibitor of tumor cell migration was found to be the anti-beta 1 monoclonal antibody 13, which inhibited the migration of human HT-1080 fibrosarcoma cells, 5637 bladder carcinoma cells, VA13 viral transformants, and HCT 116 colon carcinoma cells when fibronectin was the migration substrate. Moreover, this antibody was particularly effective in blocking cell migration on laminin, as well as migration within 3-dimensional collagen gels. It also inhibited in vitro invasiveness in a reconstituted basement membrane invasion assay (Matrigel assay) at concentrations as low as 1 microgram/ml. Integrins of the beta 1 class thus appear to play a central role in several types of migration by a variety of human tumor cell lines. Anti-alpha 5 fibronectin receptor monoclonal antibody 16 also significantly inhibited migration on fibronectin, but not on other substrates, in 3 of the 4 cell lines. Conversely, anti-alpha 2 monoclonal antibody F17 strikingly inhibited migration in 3-dimensional collagen gels, but not on other substrates, implicating the alpha 2 beta 1 integrin system in migration of tumor cells within collagenous matrices. A series of synthetic peptides previously reported to inhibit interactions of normal cells with fibronectin, laminin, and collagen were also tested as inhibitors of tumor cell migration. Peptides containing the Arg-Gly-Asp adhesive recognition signal were partially inhibitory, but with occasional exceptions, most other peptides had no effects on migration. Our results indicate the central importance of several specific beta 1 integrins in human tumor cell migration and show the effectiveness of monoclonal antibody treatment in blocking this process in vitro.
人类肿瘤细胞的迁移和侵袭过程可能涉及特定的细胞表面受体，如细胞外基质分子纤连蛋白、层粘连蛋白和胶原蛋白的受体。我们使用一组针对β1整合素家族的单克隆抗体以及一系列据报道可抑制这些蛋白质与细胞表面各种相互作用的合成肽，研究了其中几种受体的作用。发现最普遍的肿瘤细胞迁移抑制剂是抗β1单克隆抗体13，当纤连蛋白作为迁移底物时，它可抑制人HT-1080纤维肉瘤细胞、5637膀胱癌细胞、VA13病毒转化细胞和HCT 116结肠癌细胞的迁移。此外，该抗体在阻断层粘连蛋白上的细胞迁移以及三维胶原凝胶内的迁移方面特别有效。它还在重构基底膜侵袭试验（基质胶试验）中以低至1微克/毫升的浓度抑制体外侵袭性。因此，β1类整合素似乎在多种人类肿瘤细胞系的几种迁移类型中起核心作用。抗α5纤连蛋白受体单克隆抗体16在4种细胞系中的3种中也显著抑制了在纤连蛋白上的迁移，但对其他底物无抑制作用。相反，抗α2单克隆抗体F17显著抑制三维胶原凝胶中的迁移，但对其他底物无抑制作用，这表明α2β1整合素系统与肿瘤细胞在胶原基质内的迁移有关。还测试了一系列先前报道可抑制正常细胞与纤连蛋白、层粘连蛋白和胶原蛋白相互作用的合成肽作为肿瘤细胞迁移抑制剂。含有精氨酸-甘氨酸-天冬氨酸黏附识别信号的肽具有部分抑制作用，但除偶尔例外，大多数其他肽对迁移无影响。我们的结果表明几种特定的β1整合素在人类肿瘤细胞迁移中至关重要，并表明单克隆抗体治疗在体外阻断这一过程的有效性。
