Effect of bleomycin on interstitial telomeric sequences of immortalized Chinese hamster ovary cells.
The effect of the radiomimetic compound bleomycin (BLM) on interstitial telomeric sequences (ITSs) was investigated in Chinese hamster ovary (CHO) cells by using PNA-FISH with a pantelomeric probe. CHO cells were exposed to increasing concentrations of BLM and chromosomal aberrations were analyzed in the first mitosis after treatment. Cytogenetic analysis revealed that 18.1% and 9.5% of the total aberrations observed in cells exposed to BLM and harvested 18h and 3h after treatment, respectively, exhibited one or more FISH-detectable telomeric signals. Most of the chromosome breaks exhibiting telomeric signals observed in BLM-treated cells occurred in the centromeric regions of chromosomes. This observation, along with the finding of entirely labeled acentric fragments in BLM-exposed cells but not in untreated cells, shows that this antibiotic induces breakage at chromosomal sites containing ITSs. In addition, our results show that heterochromatic ITSs are involved more than expected in the formation of chromosome/chromatid breaks - and perhaps chromatid exchanges - induced by BLM, taking into account the percentage of the genome covered by telomeric sequences. On the contrary, our data strongly suggest that ITSs are not preferentially involved in the formation of dicentrics, multicentrics, centric rings, acentric fragments or chromatid deletions induced by BLM. Moreover, our results show that BLM is capable of inducing amplification and translocation of telomeric repeats, and suggest that this antibiotic produces breakage within centromeric ITSs, although chromosome regions containing these sequences are not the preferential target for BLM clastogenic action. On the other hand, our results show that BLM treatment increases the size of ITSs and that this effect is not related to the chromosomal sensitivity of the exposed cells to this compound.
Sánchez
Julieta
J
Laboratorio de Citogenética y Mutagénesis, Instituto Multidisciplinario de Biología Celular (IMBICE, CONICET La Plata-CICPBA), C.C. 403, Buenos Aires, Argentina.
Bianchi
Martha S
MS
Bolzán
Alejandro D
AD
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D001761:Bleomycin / Q000494:pharmacology*; D016466:CHO Cells; D002869:Chromosome Aberrations / Q000187:drug effects*; D006224:Cricetinae; D003412:Cricetulus; D017404:In Situ Hybridization, Fluorescence; D008938:Mitosis / Q000187:drug effects; D015335:Molecular Probes; D020135:Peptide Nucleic Acids / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid / Q000187:drug effects* / Q000235:genetics; D016615:Telomere / Q000187:drug effects* / Q000235:genetics
D000903:Antibiotics, Antineoplastic; D015335:Molecular Probes; D020135:Peptide Nucleic Acids; D001761:Bleomycin