The alpha3 laminin subunit, alpha6beta4 and alpha3beta1 integrin coordinately regulate wound healing in cultured epithelial cells and in the skin.
The alpha3 laminin subunit, alpha6beta4 and alpha3beta1 integrin coordinately regulate wound healing in cultured epithelial cells and in the skin.
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发表时间：
1999-08
影响因子：
4
通讯作者：
Lawrence E. Goldfinger;S. Hopkinson;Gregory W Dehart;Sherry S. Collawn;J. Couchman;Jonathan C. R. Jones
中科院分区：
文献类型：
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作者：
Lawrence E. Goldfinger;S. Hopkinson;Gregory W Dehart;Sherry S. Collawn;J. Couchman;Jonathan C. R. Jones
此前，我们证明了层粘连蛋白-5(LN5)α3亚单位球状结构域内的蛋白质分解处理将LN5从细胞运动诱导因子转换为能够触发半桥粒形成的蛋白质复合体，半桥粒是在上皮细胞中发现的某些细胞-基质附着部位。我们制备了一种单抗(12C4)，其表位位于α3层粘连蛋白亚单位球状结构域的羧基末端。由于蛋白水解性处理，该表位从α3亚基中丢失。抗体12C4在不能处理α3层粘连蛋白亚单位的细胞的基质中染色，但不识别MCF-10A细胞融合培养的基质，而融合培养的MCF-10A细胞有效地处理其α3层粘连蛋白链。在MCF-10A细胞的亚融合群体中，12C4仅在细胞集落的外缘染色。在这些细胞中，整合素α3beta1偶尔会与12C4抗体产生的染色共存，但整合素α6beta4不会。在受伤的MCF-10A细胞培养中，12C4抗体在覆盖伤口部位的细胞膜的最边缘，将细胞外基质染色在这些细胞下面。在人类皮肤伤口中也观察到了类似的现象，因为我们还检测到在体内将皮肤伤口上皮化的表皮细胞膜的顶端表达未经处理的α3层粘连蛋白亚单位。此外，利用Alpha3层粘连蛋白亚基和整合素功能抑制抗体，我们提供了证据，证明LN5及其两个整合素受体(Alpha6beta4和Alpha3beta1)似乎是MCF-10A细胞培养伤口愈合所必需的。基于这些结果，我们提出了损伤上皮组织愈合的模型。
Previously, we demonstrated that proteolytic processing within the globular domain of the alpha3 subunit of laminin-5 (LN5) converts LN5 from a cell motility-inducing factor to a protein complex that can trigger the formation of hemidesmosomes, certain cell-matrix attachment sites found in epithelial cells. We have prepared a monoclonal antibody (12C4) whose epitope is located toward the carboxy terminus of the globular domain of the alpha3 laminin subunit. This epitope is lost from the alpha3 subunit as a consequence of proteolytic processing. Antibody 12C4 stains throughout the matrix of cells that fail to process the alpha3 laminin subunit, but does not recognize the matrix of confluent cultures of MCF-10A cells, which efficiently process their alpha3 laminin chain. In subconfluent populations of MCF-10A cells, 12C4 only stains matrix deposited at the outer edges of cell colonies. In these cells, integrin alpha3beta1 occasionally colocalizes with the staining generated by the 12C4 antibody but alpha6beta4 integrin does not. In wounded MCF-10A cell cultures, the 12C4 antibody stains the extracellular matrix beneath those cells at the very edge of the cellular sheet that moves to cover the wound site. A similar phenomenon is observed in human skin wounds, since we also detect expression of the unprocessed alpha3 laminin subunit at the leading tip of the sheet of epidermal cells that epithelializes skin wounds in vivo. In addition, using alpha3 laminin subunit and integrin function-inhibiting antibodies, we provide evidence that LN5 and its two integrin receptors (alpha6beta4 and alpha3beta1) appear necessary for wound healing to occur in MCF-10A cell culture wounds. We propose a model for healing of wounded epithelial tissues based on these results.