Iwaki T, Iwaki A, Tateishi J, Goldman J E
Department of Neuropathology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
J Cell Biol. 1994 Jun;125(6):1385-93. doi: 10.1083/jcb.125.6.1385.
The phenotypic effects of selectively altering the levels of alpha B-crystallin in cultured glial cells were analyzed using sense and antisense approaches. Rat C6 glioma cells and human U-373MG glioma cells were transfected with a rat alpha B-crystallin sense cDNA or an antisense cDNA regulated by a Rous sarcoma virus promoter to alter cellular levels of alpha B-crystallin. The antisense strategy resulted in decreased alpha B-crystallin levels, as revealed by Western blot and immunocytochemical analyses. The reduced alpha B-crystallin expression was accompanied by alterations in cellular phenotype: (a) a reduction of cell size and/or a slender cell morphology; (b) a disorganized microfilament network; and (c) a reduction of cell adhesiveness. Like HSP27, the presence of additional alpha B-crystallin protein confers a thermoresistant phenotype to stable transfectants. Thus, alpha B-crystallin in glioma cells plays a role in their thermal resistance and may contribute to the stability of cytoskeletal organization.
采用正义和反义方法分析了在培养的神经胶质细胞中选择性改变αB-晶状体蛋白水平的表型效应。用由劳氏肉瘤病毒启动子调控的大鼠αB-晶状体蛋白正义cDNA或反义cDNA转染大鼠C6胶质瘤细胞和人U-373MG胶质瘤细胞，以改变细胞内αB-晶状体蛋白水平。如蛋白质印迹法和免疫细胞化学分析所示，反义策略导致αB-晶状体蛋白水平降低。αB-晶状体蛋白表达的降低伴随着细胞表型的改变：（a）细胞大小减小和/或细胞形态细长；（b）微丝网络紊乱；（c）细胞黏附性降低。与热休克蛋白27（HSP27）一样，额外的αB-晶状体蛋白的存在赋予稳定转染子耐热表型。因此，胶质瘤细胞中的αB-晶状体蛋白在其耐热性中起作用，并可能有助于细胞骨架组织的稳定性。
