Menter D G, Patton J T, Updyke T V, Kerbel R S, Maamer M, McIntire L V, Nicolson G L
Department of Tumor Biology, University of Texas, M. D. Anderson Cancer Center, Houston.
Cell Biophys. 1991 Apr;18(2):123-43. doi: 10.1007/BF02989810.
To resist substantial wall shear stress (WSS) exerted by flowing blood, metastatic melanoma cells can form adhesive contacts with subendothelial extracellular matrix proteins, such as fibronectin (FN). Such contacts may be stabilized by transglutaminase catalyzed-cross-linkage of cell focal adhesion proteins. We analyzed human melanoma cell adhesion under flow by decreasing the flow (WSS) of melanoma cell suspensions and allowing them to adhere to immobilized wheat germ agglutinin or FN. At the wall shear adhesion threshold (WSAT), cell adherence was rapid with no rolling. Following cell adherence, we increased the flow and determined the wall shear detachment threshold (WSDeT). Cells spread and remained adherent on immobilized FN at high WSDeTs (greater than or equal to 32.5 dynes/cm2). The high resistance of adherent cells to shear forces suggested that transglutaminase-mediated crosslinking might be involved. Transglutaminase inhibitors monodansylcadaverine and INO-3178 decreased WSAT, and at low concentrations completely inhibited tumor cell spreading and promoted detachment at low WSDeTs (0.67 dynes/cm2). In static adhesion assays, transglutaminase inhibitors decreased cell adhesion to immobilized-FN in a dose-dependent manner and prevented the formation of crosslinked 125I-FN complex that failed to enter a SDS-polyacrylamide gradient gel. The data suggest that transglutaminase-catalyzed crosslinking, particularly in the presence of WSS, may be important in stabilizing cellular adhesive contacts during adhesion to immobilized-FN.
为了抵抗流动血液施加的显著壁面剪应力（WSS），转移性黑色素瘤细胞可与内皮下细胞外基质蛋白（如纤连蛋白（FN））形成黏附接触。这种接触可通过转谷氨酰胺酶催化的细胞黏着斑蛋白交联来稳定。我们通过降低黑色素瘤细胞悬液的流速（WSS）并使其黏附于固定化的麦胚凝集素或FN，分析了流动状态下的人黑色素瘤细胞黏附情况。在壁面剪应力黏附阈值（WSAT）时，细胞黏附迅速且无滚动。细胞黏附后，我们增加流速并确定壁面剪应力脱离阈值（WSDeT）。在高WSDeT（大于或等于32.5达因/平方厘米）时，细胞在固定化FN上铺展并保持黏附。黏附细胞对剪切力的高抗性表明可能涉及转谷氨酰胺酶介导的交联。转谷氨酰胺酶抑制剂单丹磺酰尸胺和INO-3178降低了WSAT，且在低浓度时完全抑制肿瘤细胞铺展并促进在低WSDeT（0.67达因/平方厘米）时的脱离。在静态黏附试验中，转谷氨酰胺酶抑制剂以剂量依赖方式降低细胞对固定化FN的黏附，并阻止形成未能进入SDS-聚丙烯酰胺梯度凝胶的交联125I-FN复合物。数据表明，转谷氨酰胺酶催化的交联，特别是在存在WSS的情况下，可能在细胞黏附于固定化FN期间稳定细胞黏附接触方面很重要。
