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整合素在正常MDCK细胞和缺乏顶端极性的转化MDCK细胞中的表达与定位。

Integrin expression and localization in normal MDCK cells and transformed MDCK cells lacking apical polarity.

作者信息

Schoenenberger C A, Zuk A, Zinkl G M, Kendall D, Matlin K S

机构信息

Maurice E. Müller Institute, Biocenter, University of Basel, Switzerland.

出版信息

J Cell Sci. 1994 Feb;107 ( Pt 2):527-41. doi: 10.1242/jcs.107.2.527.

Abstract

Epithelial cells polarize in response to contacts with the extracellular matrix and with neighboring cells. Interactions of cells with the extracellular matrix are mediated mainly by the integrin family of receptors. To begin to understand the role of integrins in polarization, we have investigated the expression and localization of three integrin families in the polarized Madin-Darby canine kidney (MDCK) epithelial cell line and in transformed MDCK cells lacking apical polarity. We find that MDCK cells express several beta 1 integrins, including alpha 2 beta 1, alpha 3 beta 1, and an unidentified integrin designated alpha x beta 1. The beta 1 integrins are the major receptors for collagens I and IV and laminin in MDCK cells, since a blocking anti-beta 1 antibody almost totally abolishes adhesion to these proteins. They also express a vitronectin receptor tentatively identified as alpha v beta 3, and the epithelial-specific integrin alpha 6 beta 4. The latter is not a laminin receptor in MDCK cells because a function blocking anti-alpha 6 antibody has no effect on cell adhesion to laminin. All three integrin families are expressed exclusively on both the basal and lateral surfaces, as determined by immunofluorescence microscopy and surface biotinylation. Transformed MDCK cells express beta 1 integrins as well as alpha v beta 3 and alpha 6 beta 4, but show alterations in the beta 1 family. Expression of alpha x is lacking, and the relative amount of the beta 1 subunit is diminished, resulting in the accumulation of Endo-H-sensitive alpha 3. In addition, surface biotinylation and immunofluorescence indicate that significant amounts of both alpha 2 beta 1 and alpha 3 beta 1 appear on not only the basolateral but also the apical plasma membrane. These results indicate that integrins are the major receptors for the extracellular matrix in MDCK cells, and that they may affect epithelial cell polarization by mediating not only cell-substratum but also cell-cell contacts.

摘要

上皮细胞会因与细胞外基质及相邻细胞接触而发生极化。细胞与细胞外基质的相互作用主要由整合素受体家族介导。为了初步了解整合素在极化过程中的作用,我们研究了三种整合素家族在极化的犬肾上皮细胞系(MDCK)以及缺乏顶端极性的转化MDCK细胞中的表达和定位。我们发现MDCK细胞表达多种β1整合素,包括α2β1、α3β1以及一种未鉴定的整合素αxβ1。β1整合素是MDCK细胞中I型和IV型胶原蛋白以及层粘连蛋白的主要受体,因为一种阻断性抗β1抗体几乎完全消除了细胞对这些蛋白质的黏附。它们还表达一种初步鉴定为αvβ3的玻连蛋白受体,以及上皮特异性整合素α6β4。在MDCK细胞中,后者不是层粘连蛋白受体,因为一种功能阻断性抗α6抗体对细胞与层粘连蛋白的黏附没有影响。通过免疫荧光显微镜和表面生物素化测定,所有三种整合素家族仅在基底和侧面表面表达。转化的MDCK细胞表达β1整合素以及αvβ3和α6β4,但β1家族出现了改变。αx缺失,β1亚基的相对量减少,导致内切糖苷酶H敏感的α3积累。此外,表面生物素化和免疫荧光表明,大量的α2β1和α3β1不仅出现在基底外侧质膜上,还出现在顶端质膜上。这些结果表明,整合素是MDCK细胞中细胞外基质的主要受体,并且它们可能不仅通过介导细胞与基质的接触,还通过介导细胞与细胞的接触来影响上皮细胞极化。

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