ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of lidamycin (LDM) on telomerase activity in human hepatoma BEL-7402 cells under the condition of LDM inducing mitotic cell death and senescence.</p><p><b>METHODS</b>Chromatin condensation was detected by co-staining with Hoechst 33342 and PI. Cell multinucleation was observed by Giemsa staining and genomic DNA was separated by agarose gel electrophoresis. Fluorescent intensity of Rho123 was determined for mitochondrial membrane potential. MTT assay and SA-beta-gal staining were employed to analyze the senescence-like phenotype. The expression of proteins was analyzed by Western blot. Telomerase activity was assayed by telomerase PCR-ELISA.</p><p><b>RESULTS</b>Mitotic cell death occurred in LDM-treated cells characterized by unique and atypical chromatin condensation, multinucleation and increased mitochondrial membrane potential. However, no apoptotic bodies or DNA ladders were found. In addition, apoptosis-related proteins remained nearly unaltered. Senescence-like phenotype was identified by increased and elongated size of cells, growth retardation, enhanced SA-beta-gal activity and the changes of senescence-related protein expression. Telomerase activity markedly decreased (P<0.01) in LDM-treated hepatoma BEL-7402 cells.</p><p><b>CONCLUSION</b>Mitotic cell death and senescence could be triggered simultaneously or sequentially after exposure of hepatoma BEL-7402 cells to LDM. The decrease in telomerase activity may play a key role in the defective mitosis and aging morphology. Further investigation of detailed mechanism is needed.</p>
Subject(s)
Humans , Aminoglycosides , Pharmacology , Antibiotics, Antineoplastic , Pharmacology , Apoptosis , Azure Stains , Benzimidazoles , Carcinoma, Hepatocellular , Pathology , Cell Nucleus , Metabolism , Cellular Senescence , Chromatin , Metabolism , DNA, Neoplasm , Dose-Response Relationship, Drug , Enediynes , Pharmacology , Genome, Human , Genetics , Liver Neoplasms , Pathology , Membrane Potential, Mitochondrial , Mitosis , Phenotype , Propidium , Telomerase , Metabolism , Time Factors , beta-Galactosidase , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To analyze the effects of aging or advanced glycation on gene expression in the cerebrum and spleen of female C57BL/6J mice.</p><p><b>METHODS</b>The gene expression profile was determined by using cDNA expression arrays containing 588 cDNA.</p><p><b>RESULTS</b>Aging and advanced glycation resulted in differential gene expression patterns of cerebrum and spleen compared with young mice. Among the 80 genes detected in cerebrum, 43 exhibited a change in mRNA ratios with aging or treatment. Thirty-four changes (79%) were common in aged and D-galactose treated mice, whereas the cerebrum from aged and AGE-lysine treated mice showed common changes in expression of 38 genes (88%). Of the 86 genes detected in spleen, 29 (34%) displayed an age-related decrease in expression, whereas 3 (3%) displayed an increase in expression levels with aging. Eighteen genes from the detectable genes exhibited expression changes in both cerebrum and spleen of mice.</p><p><b>CONCLUSIONS</b>The gene expression profiles of D-galactose and AGE-lysine treated mice resemble those of aged mice. Use of cDNA hybridization arrays may provide a promising tool to explore the mechanism of aging at a molecular level.</p>