RAPID SPREADING AND MATURE HEMIDESMOSOME FORMATION IN HACAT KERATINOCYTES INDUCED BY INCUBATION WITH SOLUBLE LAMININ-5R
RAPID SPREADING AND MATURE HEMIDESMOSOME FORMATION IN HACAT KERATINOCYTES INDUCED BY INCUBATION WITH SOLUBLE LAMININ-5R
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DOI：
10.1111/1523-1747.ep12323451
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发表时间：
1995-10-01
影响因子：
6.5
通讯作者：
QUARANTA, V
中科院分区：
文献类型：
--
作者：
HORMIA, M;FALKMARZILLIER, J;QUARANTA, V
HaCaT细胞，一种永生化的角质形成细胞系，在804 G细胞的条件培养基存在下在塑料威尔斯孔中孵育，粘附并在不到30分钟内迅速扩散。相反，在成纤维细胞或角质形成细胞条件培养基中接种的细胞粘附较差，并在30分钟时保持圆形。抑制了细胞的快速扩散。HaCaT细胞在804G条件培养基而非对照培养基中孵育24小时后，电子显微镜显示成熟的半桥粒。在用804 G条件培养基或804 G细胞沉积基质预包被的威尔斯孔中也观察到快速扩散，但用纤连蛋白、玻连蛋白或层粘连蛋白-1预包被的孔中未观察到。804G条件培养基与抗层粘连蛋白-5r抗体的免疫印迹揭示了150，140，135，和100 kDa的多肽，相同的电泳迁移率的免疫反应性多肽在804G沉积的矩阵。我们的研究结果表明，除了层粘连蛋白-5r在一个可溶性的形式是足以促进角质形成细胞中的快速蔓延和半桥粒组装。可溶性层粘连蛋白-5r的作用机制可能包括有效的表面"引发"细胞粘附。可溶性层粘连蛋白-5r可能在表皮的形态发生和修复中具有生理作用，并且可能用于治疗应用。
HaCaT cells, an immortalized keratinocyte line, incubated in plastic wells in the presence of conditioned medium from 804G cells adhered and spread rapidly in less than 30 min. In contrast, cells plated in fibroblast or keratinocyte conditioned medium adhered poorly and remained rounded at 30 min. Immunodepletion of 804G conditioned medium with polyclonal antisera to laminin-5r, but not control antisera, abolished rapid cell spreading. Electron microscopy of HaCaT cells spread by incubation in 804G conditioned medium, but not control medium, revealed mature hemidesmosomes after 24 h. Rapid spreading was also observed in wells precoated with 804G conditioned medium or 804G cell-deposited matrix, but not with fibronectin, vitronectin, or laminin-1. Immunoblotting of 804G conditioned medium with anti-laminin-5r antibodies unveiled polypeptides of 150, 140, 135, and 100 kDa, identical by electrophoretic mobility to immunoreactive polypeptides in 804G deposited matrix. Our results suggest that addition of laminin-5r in a soluble form is sufficient to promote rapid spreading and hemidesmosome assembly in keratinocytes. The mechanism of soluble laminin-5r action may include efficient surface ''priming'' for cell adhesion. Soluble laminin-5r may have a physiologic role in morphogenesis and repair of the epidermis and may be of use for therapeutic applications.