Inhibition of laminin alpha 1-chain expression leads to alteration of basement membrane assembly and cell differentiation.
Inhibition of laminin alpha 1-chain expression leads to alteration of basement membrane assembly and cell differentiation.
复制标题
抑制层粘连蛋白α1-链表达会导致基底膜组装和细胞分化的改变。
DOI：
10.1083/jcb.133.2.417
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发表时间：
1996-04
影响因子：
7.8
通讯作者：
Simon-Assmann, P
中科院分区：
文献类型：
--
作者：
De Arcangelis, A;Neuville, P;Boukamel, R;Lefebvre, O;Kedinger, M;Simon-Assmann, P
层粘连蛋白-1（基底膜的主要成分）的组成性α 1链的表达在发育和分化过程中受到显著调节。我们设计了一种反义RNA策略来分析α 1链在层粘连蛋白组装、基底膜形成和细胞分化中的直接参与。我们报道了α 1链表达的缺失，这是由用包含以反义方向插入的α 1链的cDNA片段的真核表达载体稳定转染人结肠癌Caco 2细胞引起的，导致（a）层粘连蛋白-1的另外两个组成链，β 1/γ 1链，（B）当Caco 2缺陷型细胞在成纤维细胞上培养时，缺乏基底膜组装，通过缺乏IV型胶原和巢蛋白沉积来评估，和（c）细胞结构极性的变化，伴随着顶端消化酶蔗糖酶-异麦芽糖酶的抑制。结果表明，α 1链是分泌层粘连蛋白-1和组装基底膜网络所必需的。此外，层粘连蛋白α 1链基因的表达可能是决定细胞分化的调节元件。
The expression of the constituent alpha 1 chain of laminin-1, a major component of basement membranes, is markedly regulated during development and differentiation. We have designed an antisense RNA strategy to analyze the direct involvement of the alpha 1 chain in laminin assembly, basement membrane formation, and cell differentiation. We report that the absence of alpha 1-chain expression, resulting from the stable transfection of the human colonic cancer Caco2 cells with an eukaryotic expression vector comprising a cDNA fragment of the alpha 1 chain inserted in an antisense orientation, led to (a) an incorrect secretion of the two other constituent chains of laminin-1, the beta 1/gamma 1 chains, (b) the lack of basement membrane assembly when Caco2-deficient cells were cultured on top of fibroblasts, assessed by the absence of collagen IV and nidogen deposition, and (c) changes in the structural polarity of cells accompanied by the inhibition of an apical digestive enzyme, sucrase-isomaltase. The results demonstrate that the alpha 1 chain is required for secretion of laminin-1 and for the assembly of basement membrane network. Furthermore, expression of the laminin alpha 1-chain gene may be a regulatory element in determining cell differentiation.