Mutagenesis, Vol. 16, No. 5, 369-375,
September 2001
© 2001 UK Environmental Mutagen Society/Oxford University Press
Evaluation of the genotoxic effects of the boron neutron capture reaction in human melanoma cells using the cytokinesis block micronucleus assay
1 Department of Genetics, Faculty of Medical Sciences, New University of Lisbon, R. da Junqueira 96, P 1349-008 Lisbon, 2 Faculty of Pharmacy, University of Lisbon, Lisbon, 3 CFEC, Faculty of Medicine, University of Lisbon, Lisbon, 4 University Lusófona, Lisbon, and 5 Nuclear and Technological Institute, Portuguese Research Reactor, Sacavém, Lisbon, Portugal
The present work reports on the genotoxicity of the boron neutron capture (BNC) reaction in human metastatic melanoma cells (A2058) assessed by the cytokinesis block micronucleus assay (CBMN) using p-borono-L-phenylalanine (BPA) as the boron delivery agent. Different concentrations of BPA (0.48, 1.2 and 2.4 mM) and different fluences of thermal neutrons were studied. Substantial genotoxic potential of 
and lithium particles generated inside or near the malignant cell by the BNC reaction was observed in a dose–response manner as measured by the frequency of micronucleated binucleated melanoma cells and by the number of micronuclei (MN) per binucleated cell. The distribution of the number of MN per micronucleated binucleated cell was also studied. The BNC reaction clearly modifies this distribution, increasing the frequency of micronucleated cells with 2 and, especially, 
3 MN and conversely decreasing the frequency of micronucleated cells with 1 MN. A decrease in cell proliferation was also observed which correlated with MN formation. A discrete genotoxic and anti-proliferative contribution from both thermal neutron irradiation and BPA was observed and should be considered secondary. Additionally, V79 Chinese hamster cells (chromosomal aberrations assay) and human lymphocytes (CBMN assay) incubated with different concentrations of BPA alone did not show any evidence of genotoxicity. The presented results reinforce the usefulness of the CBMN assay as an alternative method for assessment of the deleterious effects induced by high LET radiation produced by the BNC reaction in human melanoma cells.
6 To whom correspondence should be addressed. Fax: +351 21 3622018; Email: rueff.gene{at}fcm.unl.pt
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