Volume 15, Issue 1 p. 221-229
Research Communication
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Lens epithelial cells derived from αB-crystallin knockout mice demonstrate hyperproliferation and genomic instability

U.P. ANDLEY

Corresponding Author

U.P. ANDLEY

Departments of Ophthalmology and Visual Sciences, Washington University School of Medicine, St. Louis, Missouri, 63110 USA

Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri, 63110 USA

Correspondence: Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, 660 S. Euclid Ave., Campus Box 8096, St. Louis, MO 63110, USA. E-mail: [email protected]Search for more papers by this author
Z. SONG

Z. SONG

Departments of Ophthalmology and Visual Sciences, Washington University School of Medicine, St. Louis, Missouri, 63110 USA

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E.F. WAWROUSEK

E.F. WAWROUSEK

National Eye Institute, National Institutes of Health, Bethesda, Maryland, 20892 USA

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J.P. BRADY

J.P. BRADY

National Eye Institute, National Institutes of Health, Bethesda, Maryland, 20892 USA

MetaMorphix, Inc., Baltimore, MD, 21227 USA

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S. BASSNETT

S. BASSNETT

Departments of Ophthalmology and Visual Sciences, Washington University School of Medicine, St. Louis, Missouri, 63110 USA

Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri, 63110 USA

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T.P. FLEMING

T.P. FLEMING

Genetics, Washington University School of Medicine, St. Louis, Missouri, 63110 USA

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First published: 01 January 2001
Citations: 50

ABSTRACT

B-crystallin is a member of the small heat shock protein family and can act as a molecular chaperone preventing the in vitro aggregation of other proteins denatured by heat or other stress conditions. Expression of αB-crystallin increases in cells exposed to stress and enhanced in tumors of neuroectodermal origin and in many neurodegenerative diseases. In the present study, we examined the properties of lens epithelial cells derived from mice in which the αB-crystallin gene had been knocked out. Primary rodent cells immortalize spontaneously in tissue culture with a frequency of 10 –5 to 10 –6. Primary lens epithelial cells derived from aB-crystaUin–/– mice produced hyperproliferative clones at a frequency of 7.6 × 10–2, four orders of magnitude greater than predicted by spontaneous immortalization (1). Hyperproliferative αB-crys-tallin–/– cells were shown to be truly immortal since they have been passaged for more than 100 population doublings without any diminution in growth potential. In striking contrast to the wild-type cells, which were diploid, the αB-crystallin–/– cultures had a high proportion of tetraploid and higher ploidy cells, indicating that the loss of αB-crystallin is associated with an increase in genomic instability. Further evidence of genomic instability of αB-crystallin–/– cells was observed when primary cultures were infected with Ad12– SV40 hybrid virus. In striking contrast to wild-type cells, αB-crystallin–/– cells expressing SV40 T antigen exhibited a widespread cytocidal response 2 to 3 days after attaining confluence, indicating that SV40 T antigen enhanced the intrinsic genomic instability of αB-crystallin–/– lens epithelial cells. These observations suggest that the widely distributed molecular chaperone αB-crystallin may play an important nuclear role in maintaining genomic integrity.—Andley, U. P., Song, Z., Wawrousek, E. F., Brady, J. P., Bassnett, S., Fleming, T. P. Lens epithelial cells derived from αB-crystallin knockout mice demonstrate hyperproliferation and genomic instability. FASEB J. 15, 221–229 (2001)