Sense and Antisense Modification of Glial αB-Crystallin Production Results in Alterations of Stress Fiber Formation and Thermoresistance

• Published in: The Journal of cell biology Vol. 125; no. 6; pp. 1385 - 1393
• Main Authors: Iwaki, Toru, Iwaki, Akiko, Tateishi, Jun, Goldman, James E.
• Format: Journal Article
• Language: English
• Published: New York, NY Rockefeller University Press 01-06-1994; The Rockefeller University Press
• Subjects: Animals; Antiserum; Biological and medical sciences; Cell Adhesion - genetics; Cell Line; Cell lines; Cells; Complementary DNA; Crystallins; Crystallins - biosynthesis; Crystallins - genetics; Cytoskeleton - physiology; DNA, Antisense - genetics; DNA, Antisense - pharmacology; DNA, Complementary - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Neoplastic; Glioma; Heat-Shock Proteins - biosynthesis; Heat-Shock Proteins - genetics; Hot Temperature - adverse effects; Humans; Messenger RNA; Microfilaments; Miscellaneous; Molecular and cellular biology; Molecular genetics; Neuroglia; Neuroglia - cytology; Neuroglia - drug effects; Neuroglia - physiology; Neuroglia - ultrastructure; Rats; Stress fibers; Transfection; Transformation, Genetic; Tumor Cells, Cultured; Resistance; Vertebrata; Glial cell; Temperature; Cell line; Mammalia; Crystallin; Rat; Morphology; Rodentia; Neuroglía; Gene expression
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.125.6.1385

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.