The phenotypic effects of selectively altering the levels of αB- 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 αB-crystallin sense cDNA or an antisense cDNA regulated by a Rous sarcoma virus promoter to alter cellular levels of αB- crystallin. The antisense strategy resulted in decreased αB-crystallin levels, as revealed by Western blot and immunocytochemical analyses. The reduced αB-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 αB-crystallin protein confers a thermoresistant phenotype to stable transfectants. Thus, αB- crystallin in glioma cells plays a role in their thermal resistance and may contribute to the stability of cytoskeletal organization.
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