Sense and antisense modification of glial alpha B-crystallin production results in alterations of stress fiber formation and thermoresistance.
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.
Iwaki
T
T
Department of Neuropathology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Iwaki
A
A
Tateishi
J
J
Goldman
J E
JE
D000818:Animals; D002448:Cell Adhesion / Q000235:genetics; D002460:Cell Line; D003459:Crystallins / Q000096:biosynthesis* / Q000235:genetics; D003599:Cytoskeleton / Q000502:physiology; D016373:DNA, Antisense / Q000235:genetics / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006358:Hot Temperature / Q000009:adverse effects; D006801:Humans; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D051381:Rats; D014162:Transfection; D014170:Transformation, Genetic; D014407:Tumor Cells, Cultured
D003459:Crystallins; D016373:DNA, Antisense; D018076:DNA, Complementary; D006360:Heat-Shock Proteins