stilbenes and Chromosome-Deletion

In recent years, a great interest has emerged in resveratrol (RSV) activity in the prevention of various pathologies including cancer. We recently showed that RSV is able to interfere with topoisomerase II-α (TOPO2) activity in cancer cells, thus inducing a delay in S-phase progression with concomitant phosphorylation of the histone H2AX. TOPO2 is mainly active in proliferating cells and is involved in the resolution of supercoiled DNA and chromosome segregation during mitosis. Here, we studied the effects of RSV in CHO-K1 cells concerning to chromosome damage and segregation as a consequence of TOPO2 inhibition. We show an increase in micronuclei and in polyploid and endoreduplicated cells due to incorrect chromosome segregation. Furthermore, since incomplete segregation can also affect the normal distribution of mitotic figures, we checked mitosis progression showing an increase in metaphase in relation to ana-telophase after RSV treatment. On the whole, our data show that RSV affects chromosome stability and segregation in proliferating cells, probably interfering with TOPO2 activity.

Topics: Animals; Antigens, Neoplasm; Antineoplastic Agents, Phytogenic; CHO Cells; Chromosome Deletion; Chromosome Segregation; Cricetinae; DNA Damage; DNA Topoisomerases, Type II; DNA-Binding Proteins; Enzyme Activation; Enzyme Inhibitors; Humans; Micronuclei, Chromosome-Defective; Mitosis; Polyploidy; Resveratrol; Stilbenes

Glutathione S-transferase (GST) class Mu activity was determined in 145 unrelated hospital patients in Berlin by measuring their conjugation activity towards the specific substrate trans-stilbene oxide (TSO) with two substrate concentrations (50 and 250 microM) in homogenates prepared from lymphocytes. Eighty individuals (55.2%) had an activity lower than 10 pmol/min/10(6) lymphocytes and were classified as GST class Mu deficient. In 142 of 145 cases, phenotype was confirmed by the results of a genotyping procedure using the polymerase chain reaction technique. Two fragments of 273 and about 650 bp including one and two introns, respectively, could always be amplified from genomic DNA in individuals with high GST class Mu activity and could not be amplified in persons with impaired glutathione-TSO conjugation activity. This indicates that persons with low activity carry a large deletion mutation within the GST class Mu gene. The enzymatically determined antimode between low and high activity determined as 10 pmol/min/1 million lymphocytes in the assay with 50 microM TSO could be clearly confirmed by genotyping.

Topics: Adult; Aged; Aged, 80 and over; Base Sequence; Berlin; Chromosome Deletion; Female; Germany; Glutathione; Glutathione Transferase; Humans; Isoenzymes; Lung Neoplasms; Lymphocytes; Male; Middle Aged; Molecular Sequence Data; Mutation; Phenotype; Polymerase Chain Reaction; Stilbenes

Glutathione transferase (GT; EC 2.5.1.18) mRNA levels were measured in human liver samples by using mouse and human cDNA clones that encode class-mu and class-alpha GT. Although all the RNA samples examined contained class-alpha GT mRNA, class-mu GT mRNA was found only in individuals whose peripheral leukocytes expressed GT activity on the substrate trans-stilbene oxide. The mouse class-mu cDNA clone was used to identify a human class-mu GT cDNA clone, lambda GTH411. The amino acid sequence of the GT encoded by lambda GTH411 is identical with the 23 residues determined for the human liver GT-mu isoenzyme and shares 76-81% identity with mouse and rat class-mu GT isoenzymes. The mouse and human class-mu GT cDNA inserts hybridize with multiple BamHI and EcoRI restriction fragments in the human genome. One of these hybridizing fragments is missing in the DNA of individuals who lack GT activity on trans-stilbene oxide. Hybridizations with nonoverlapping subfragments of lambda GTH411 suggest that there are at least three class-mu genes in the human genome. One of these genes appears to be deleted in individuals lacking GT activity on trans-stilbene oxide.

Topics: Amino Acid Sequence; Animals; Base Sequence; Chromosome Deletion; Glutathione Transferase; Humans; Isoenzymes; Liver; Molecular Sequence Data; Rats; RNA, Messenger; Stilbenes