Hyperproliferation and p53 status of lens epithelial cells derived from alphaB-crystallin knockout mice

التفاصيل البيبلوغرافية
العنوان: Hyperproliferation and p53 status of lens epithelial cells derived from alphaB-crystallin knockout mice
المؤلفون: Fang Bai, Timothy P. Fleming, Jing Hua Xi, Eric F. Wawrousek, Usha P. Andley
المصدر: The Journal of biological chemistry. 278(38)
سنة النشر: 2003
مصطلحات موضوعية: Genome instability, DNA, Complementary, Time Factors, Cell division, Binucleated cells, Blotting, Western, Centromere, Immunoblotting, Mitosis, Apoptosis, Mice, Transgenic, Biology, Biochemistry, Retinoblastoma Protein, Mice, Lens, Crystalline, medicine, Animals, Muscle, Skeletal, Molecular Biology, Cells, Cultured, In Situ Hybridization, Fluorescence, Mice, Knockout, medicine.diagnostic_test, Cell Death, Cell Cycle, Wild type, alpha-Crystallin B Chain, Epithelial Cells, Cell Biology, DNA, Cell biology, Phenotype, Microscopy, Fluorescence, Gamma Rays, biology.protein, Antibody, Tumor Suppressor Protein p53, Cytokinesis, Cell Division, Fluorescence in situ hybridization
الوصف: alphaB-Crystallin, a major protein of lens fiber cells, is a stress-induced chaperone expressed at low levels in the lens epithelium and numerous other tissues, and its expression is enhanced in certain pathological conditions. However, the function of alphaB in these tissues is not known. Lenses of alphaB-/- mice develop degeneration of specific skeletal muscles but do not develop cataracts. Recent work in our laboratory indicates that primary cultures of alphaB-/- lens epithelial cells demonstrate genomic instability and undergo hyperproliferation at a frequency 4 orders of magnitude greater than that predicted by spontaneous immortalization of rodent cells. We now demonstrate that the hyperproliferative alphaB-/- lens epithelial cells undergo phenotypic changes that include the appearance of the p53 protein as shown by immunoblot analysis. Sequence analysis showed a lack of mutations in the p53 coding region of hyperproliferative alphaB-/- cells. However, the reentry of hyperproliferative alphaB-/- cells into S phase and mitosis after DNA damage by gamma-irradiation were consistent with impaired p53 checkpoint function in these cells. The results demonstrate that expression of functionally impaired p53 is one of the factors that promote immortalization of lens epithelial cells derived from alphaB-/- mice. Fluorescence in situ hybridization using probes prepared from centromere-specific mouse P1 clones of chromosomes 1 and 9 demonstrated that the hyperproliferative alphaB-/- cells were 30% diploid and 70% tetraploid, whereas wild type cells were 83% diploid. Further evidence of genomic instability was obtained when the hyperproliferative alphaB-/- cells were labeled with anti-beta-tubulin antibodies. Examination of the hyperproliferative alphaB-/- mitotic profiles revealed the presence of cells that failed to round up for mitosis, or arrested in cytokinesis, and binucleated cells in which nuclear division had occurred without cell division. These results suggest that the stress protein and molecular chaperone alphaB-crystallin protects cells from acquiring impaired p53 protein and genomic instability.
تدمد: 0021-9258
الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::df13770a63f82da1f446d44fdccc77eaTest
https://pubmed.ncbi.nlm.nih.gov/12826669Test
حقوق: OPEN
رقم الانضمام: edsair.doi.dedup.....df13770a63f82da1f446d44fdccc77ea
قاعدة البيانات: OpenAIRE