Koivisto L, Heino J, Häkkinen L, Larjava H
Institute of Dentistry, University of Turku, Finland.
Biochem J. 1994 Jun 15;300 ( Pt 3)(Pt 3):771-9. doi: 10.1042/bj3000771.
A large pool of precursor beta 1-integrin subunits is frequently found intracellularly. During malignant transformation this pool often disappears. Concomitantly, integrin-mediated cell-adhesion functions are disturbed, even though no change in the number of beta 1-integrin receptors on the cell surface can be observed. Here, we have studied the role of an intracellular pre-beta 1-integrin pool by transfecting human MG-63 osteosarcoma cells with plasmid construction producing an antisense RNA for the beta 1-integrin subunit. Stable cell clones expressing beta 1-integrin antisense RNA were shown to have a reduced intracellular pool of pre-beta 1-integrin subunits. In the antisense-transfected cells, the synthesis of the beta 1-integrin chain was reduced by 65% compared with non-transfected or vector-transfected MG-63 cells. The decreased synthesis of the beta 1-integrin chain was associated with accelerated maturation of the beta 1-integrin chain (half-maturation time about 5 h in antisense-transfected cells compared with about 10.5 h in control cells), whereas maturation of the alpha-integrin chain slowed down. The amount of beta 1-integrins on the cell surface, however, remained unaltered. Cell clones with the largest decrease in the relative amount of the pre-beta 1-integrin subunit also showed altered integrin function. They were found to synthesize fibronectin, but were unable to assemble it into a fibronectin matrix on the cell surface. Thus we conclude that the repression of biosynthesis of the beta 1-integrin chain leads to alterations in receptor maturation and may be connected with altered receptor function.
细胞内经常发现大量的前体β1整合素亚基。在恶性转化过程中,这一亚基库常常消失。与此同时,尽管在细胞表面未观察到β1整合素受体数量的变化,但整合素介导的细胞黏附功能却受到了干扰。在此,我们通过用产生β1整合素亚基反义RNA的质粒构建体转染人MG-63骨肉瘤细胞,研究了细胞内前β1整合素亚基库的作用。结果显示,表达β1整合素反义RNA的稳定细胞克隆的前β1整合素亚基细胞内库减少。在反义转染细胞中,与未转染或载体转染的MG-63细胞相比,β1整合素链的合成减少了65%。β1整合素链合成的减少与β1整合素链成熟加速有关(反义转染细胞中的半成熟时间约为5小时,而对照细胞中约为10.5小时),而α整合素链的成熟则减慢。然而,细胞表面β1整合素的数量保持不变。前β1整合素亚基相对量减少最多的细胞克隆也显示出整合素功能改变。它们能合成纤连蛋白,但无法将其组装成细胞表面的纤连蛋白基质。因此我们得出结论,β1整合素链生物合成的抑制导致受体成熟的改变,并可能与受体功能的改变有关。