Marshall J F, Rutherford D C, McCartney A C, Mitjans F, Goodman S L, Hart I R
Richard Dimbleby Department of Cancer Research/ICRF Laboratory, St Thomas' Hospital, London, UK.
J Cell Sci. 1995 Mar;108 ( Pt 3):1227-38. doi: 10.1242/jcs.108.3.1227.
We have shown previously that VUP was the only line out of ten human melanoma lines that failed to express the vitronectin receptor alpha v beta 3, but instead expressed alpha v beta 1. Levels of alpha v beta 1 expression were low on parental VUP cells so that iterative sorting by FACS, using an anti-alpha v antibody (13C2), was utilised to derive sublines with 8- to 10-fold higher amounts of cell surface alpha v beta 1. There was little difference between low (V-) and high (V+) alpha v beta 1-expressing sublines with regard to adherence to collagen type I, collagen type IV or laminin substrata. However, adherence to vitronectin and fibrinogen correlated closely with alpha v beta 1 expression (35-42% adhesion for V(+) lines versus 6-8% adhesion for V- lines on vitronectin, for example). Utilising a high alpha v beta 1-expressing subline (V + B2) we have shown that binding to vitronectin and fibrinogen was inhibited specifically by function-blocking antibodies to alpha v (17E6 and 14D9) and beta 1 (A11B2). V(+) sublines spread more compared with V(-) sublines on both vitronectin and fibronectin. However, neither alpha 5- nor alpha v-blocking antibodies had any effect on attachment or spreading of V + B2 on fibronectin whereas the combination of alpha 5 (PID6)- and alpha v(17E6)-blocking antibodies abrogated binding to fibronectin almost completely. This is the first report of an alpha v beta 1 integrin able to recognize vitronectin and fibrinogen, and also cooperate with alpha 5 beta 1 to mediate attachment to and spreading on fibronectin.
我们之前已经表明,VUP是十条人类黑色素瘤细胞系中唯一不表达玻连蛋白受体αvβ3,而是表达αvβ1的细胞系。亲代VUP细胞上αvβ1的表达水平较低,因此利用抗αv抗体(13C2)通过荧光激活细胞分选术(FACS)进行反复分选,以获得细胞表面αvβ1含量高8至10倍的亚系。低表达(V-)和高表达(V+)αvβ1的亚系在黏附于I型胶原、IV型胶原或层粘连蛋白基质方面几乎没有差异。然而,黏附于玻连蛋白和纤维蛋白原与αvβ1的表达密切相关(例如,在玻连蛋白上,V(+)细胞系的黏附率为35 - 42%,而V-细胞系为6 - 8%)。利用一个高表达αvβ1的亚系(V + B2),我们已经表明,与玻连蛋白和纤维蛋白原的结合被针对αv(17E6和14D9)和β1(A11B2)的功能阻断抗体特异性抑制。与V(-)亚系相比,V(+)亚系在玻连蛋白和纤连蛋白上的铺展更多。然而,α5阻断抗体和αv阻断抗体对V + B2在纤连蛋白上的附着或铺展均无任何影响,而α5(PID6)阻断抗体和αv(17E6)阻断抗体的组合几乎完全消除了与纤连蛋白的结合。这是关于一种能够识别玻连蛋白和纤维蛋白原,并且还能与α5β1协同介导在纤连蛋白上附着和铺展的αvβ1整合素的首次报道。