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在表现出不同形态和致瘤特性的小鼠杂交种中的基因表达。

Gene expression in murine hybrids exhibiting different morphologies and tumorigenic properties.

作者信息

Bravo R, Fey S J, Celis J E

出版信息

Carcinogenesis. 1981;2(8):769-82. doi: 10.1093/carcin/2.8.769.

Abstract

[35S]Methionine labelled polypeptides from mouse CLID, hamster ovary cells (CHO, and 7 derived somatic cell hybrids which have segregated CHO chromosomes, were analysed by means of high resolution two-dimensional gel electrophoresis under conditions in which the position of 600 polypeptides could be reproducibly assessed. As judged from the two-dimensional gel electrophoretic patterns (isoelectric focussing (IEF) and non-equilibrium pH gradient electrophoresis (NEPHGE)) gene expression in all the hybrids resembled the mouse CLID parent and with only one exception they all expressed different numbers and intensities of CHO specific polypeptides. Even though some of the hybrids expressed as much as 50% of the total number of CHO specific polypeptides that could be clearly differentiated from those of the mouse parent we failed to find a direct correlation between the expression of any given CHO polypeptide and the morphological or tumorigenic properties of the hybrids. Most CHO specific polypeptides, however, were expressed at lower levels in the hybrids as compared to the parent CHO cells, a fact that may be due to the chromosomal constitution of the hybrids, regulation or both. Similarly, the quantitation of the major cytoskeletal polypeptides present in the hybrids, such as alpha-and beta-tubulin, vimentin, total actin and 3 polypeptides (IEF 12, 24 and 31) present in intermediate filament enriched cytoskeletons, indicate that changes in the relative proportion of any of these proteins is not sufficient to account for the morphology, actin microfilament pattern or tumorigenicity of the hybrids. Co-expression of cytoskeletal proteins in the hybrids could only be demonstrated in the case of the related mouse IEF 24 and hamster IEF 7 polypeptides. In all other cases the cytoskeletal polypeptides co-migrated and presented similar one-dimensional peptide maps. Some principles are emerging concerning the possibility of using somatic cell hybridization in combination with two-dimensional gel electrophoresis to locate genes coding for particular polypeptides on a given chromosome.

摘要

用[35S]甲硫氨酸标记来自小鼠CLID、仓鼠卵巢细胞(CHO)以及7个已分离出CHO染色体的体细胞杂种的多肽,在可重复评估600种多肽位置的条件下,通过高分辨率二维凝胶电泳进行分析。从二维凝胶电泳图谱(等电聚焦(IEF)和非平衡pH梯度电泳(NEPHGE))判断,所有杂种中的基因表达都类似于小鼠CLID亲本,除了一个例外,它们都表达了不同数量和强度的CHO特异性多肽。尽管有些杂种表达的CHO特异性多肽总数高达可与小鼠亲本明显区分的50%,但我们未能发现任何特定CHO多肽的表达与杂种的形态或致瘤特性之间存在直接关联。然而,与亲本CHO细胞相比,大多数CHO特异性多肽在杂种中的表达水平较低,这一事实可能归因于杂种的染色体组成、调控或两者兼而有之。同样,对杂种中存在的主要细胞骨架多肽进行定量分析,如α-和β-微管蛋白、波形蛋白、总肌动蛋白以及存在于富含中间丝的细胞骨架中的3种多肽(IEF 12、24和31),结果表明这些蛋白质中任何一种相对比例的变化都不足以解释杂种的形态、肌动蛋白微丝模式或致瘤性。细胞骨架蛋白在杂种中的共表达仅在相关的小鼠IEF 24和仓鼠IEF 7多肽的情况下得到证实。在所有其他情况下,细胞骨架多肽共迁移并呈现相似的一维肽图。关于将体细胞杂交与二维凝胶电泳结合使用来定位特定染色体上编码特定多肽的基因的可能性,一些原则正在显现。

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