Ducros E, Berthaut A, Mirshahi P, Lemarchand S, Soria J, Legeais J M, Mirshahi M
UMRS 736 INSERM - Université Pierre et Marie Curie (Paris 6), Faculté de Médecine Paris VI, Paris, France.
Curr Eye Res. 2007 Jun;32(6):481-90. doi: 10.1080/02713680701411269.
The fibulins are a family of extracellular matrix (ECM) molecules that regulate the organ shape along with other growth factors and stromal cells. We report here the in vitro expression of ECM proteins fibulin-1 and fibulin-2 by human corneal fibroblasts. The ability of fibulin-1 to modulate cell motility was investigated.
Fibulin-1 and fibulin-2 mRNA and proteins expression were analyzed in primary and immortalized human corneal fibroblasts (CHN) respectively by gene array, RT-PCR, and immunocytochemistry. The motility and adhesion of the cells transfected with fibulin-1 siRNA were analyzed on tissue culture polystyrene coated with Matrigel or ECM secreted by those fibroblasts.
(1) The microarray analysis shows the expression of fibulin-1, fibulin-2, and their binding partners (i.e., fibronectin, nidogen-1, aggrecan, fibrilin-1, endostatin, and laminin alpha-2 chain). Interestingly, a matrix metalloprotease, ADAMTS-1, for which fibulin-1 acts as a cofactor, was also detected in CHN. (2) The synthesis by CHN of fibulin-1 and 2 mRNA and proteins was confirmed respectively by RT-PCR and immunocytochemistry. (3) Transfection of CHN by fibulin-1 siRNA has no effect on cell adhesion but increases cell migration compared with that of the control cells. This observation suggests an important role of fibulin-1 on cell motility.
The expression of fibulins and that of their binding partners by human corneal fibroblasts indicate the important role of these proteins in the organization of supramolecular ECM structures of cornea. The variation of their expression and the structural changes of fibulins remain to be determined in corneal pathology.
纤维连接蛋白是一类细胞外基质（ECM）分子，与其他生长因子和基质细胞共同调节器官形态。我们在此报告人角膜成纤维细胞在体外表达ECM蛋白纤维连接蛋白-1和纤维连接蛋白-2的情况。研究了纤维连接蛋白-1调节细胞运动的能力。
分别通过基因芯片、逆转录-聚合酶链反应（RT-PCR）和免疫细胞化学分析原代和永生化人角膜成纤维细胞（CHN）中纤维连接蛋白-1和纤维连接蛋白-2的信使核糖核酸（mRNA）及蛋白表达。在涂有基质胶或这些成纤维细胞分泌的ECM的组织培养聚苯乙烯上分析转染了纤维连接蛋白-1小干扰RNA（siRNA）的细胞的运动性和黏附性。
（1）微阵列分析显示了纤维连接蛋白-1、纤维连接蛋白-2及其结合伴侣（即纤连蛋白、巢蛋白-1、聚集蛋白聚糖、原纤蛋白-1、内皮抑素和层粘连蛋白α-2链）的表达。有趣的是，在CHN中还检测到一种基质金属蛋白酶ADAMTS-1，纤维连接蛋白-1作为其辅因子发挥作用。（2）通过RT-PCR和免疫细胞化学分别证实了CHN合成纤维连接蛋白-1和2的mRNA及蛋白。（3）与对照细胞相比，用纤维连接蛋白-1 siRNA转染CHN对细胞黏附没有影响，但增加了细胞迁移。这一观察结果表明纤维连接蛋白-1在细胞运动中起重要作用。
人角膜成纤维细胞表达纤维连接蛋白及其结合伴侣表明这些蛋白在角膜超分子ECM结构组织中起重要作用。它们的表达变化和纤维连接蛋白的结构变化在角膜病理学中仍有待确定。
