Judware R, Culp L A
Department of Molecular Biology and Microbiology, Case Western Reserve University, School of Medicine, Cleveland, OH 44106, USA.
Clin Exp Metastasis. 1997 May;15(3):228-38. doi: 10.1023/a:1018417330479.
Alterations in adhesion to the extracellular matrix mediated by integrin receptors are commonly observed in a wide variety of transformed/tumor classes. Reductions in the expression of several integrin subunits have been documented in human neuroblastoma cell lines that over-express the neuroblastoma-associated oncogene N-myc. Neuroblastoma cells transfected with a cDNA encoding N-myc on a high-expression plasmid exhibit greatly reduced levels of alpha2, alpha3 and beta1 integrin subunits with concomitant rounding of cells on substrata. In the current studies, we examined whether integrin downregulation by N-myc is cell-type specific by transfecting a human N-myc cDNA into Saos-2 human osteosarcoma cells and evaluating integrin expression. Several N-myc-expressing cell lines were isolated which exhibit reduced levels of beta1 integrin subunit protein and significant alteration in cell morphology - these cell lines resemble N-myc-over-expressing neuroblastoma cells. In addition to reduced beta1 subunit levels, the osteosarcoma-derived N-myc transfectants exhibit little or no alpha3beta1 integrin complexes, either intracellular or at the cell surface. Finally, reduced amounts of alpha3 integrin subunit in these cell lines occur at the level of alpha3 integrin mRNA, although post-transcriptional mechanisms may also be involved, particularly with inability of pre-beta1 protein to mature. These results confirm our previous studies demonstrating integrin downregulation by an N-myc-dependent process and, in addition, demonstrate lack of cell-type specificity in the action of N-myc on integrin extracellular matrix receptor expression when comparing neural precursor (neuroblastoma) cells with connective tissue (osteosarcoma) cells.
整合素受体介导的与细胞外基质黏附的改变在多种转化/肿瘤类型中普遍可见。在过表达神经母细胞瘤相关癌基因N - myc的人神经母细胞瘤细胞系中,已记录到几种整合素亚基的表达降低。用高表达质粒上编码N - myc的cDNA转染的神经母细胞瘤细胞,其α2、α3和β1整合素亚基水平大幅降低,同时细胞在基质上变圆。在当前研究中,我们通过将人N - myc cDNA转染到Saos - 2人骨肉瘤细胞中并评估整合素表达,来检验N - myc对整合素的下调是否具有细胞类型特异性。分离出了几个表达N - myc的细胞系,这些细胞系显示β1整合素亚基蛋白水平降低且细胞形态有显著改变——这些细胞系类似于过表达N - myc的神经母细胞瘤细胞。除了β1亚基水平降低外,源自骨肉瘤的N - myc转染细胞在细胞内或细胞表面几乎没有α3β1整合素复合物。最后,这些细胞系中α3整合素亚基数量的减少发生在α3整合素mRNA水平,尽管转录后机制可能也起作用,特别是前β1蛋白无法成熟时。这些结果证实了我们之前的研究,即通过N - myc依赖的过程下调整合素,此外,在比较神经前体细胞(神经母细胞瘤)和结缔组织细胞(骨肉瘤)时,还证明了N - myc对整合素细胞外基质受体表达的作用缺乏细胞类型特异性。