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Appl Radiat Isot. 2009 Jul;67(7-8 Suppl):S325-7. doi: 10.1016/j.apradiso.2009.03.072. Epub 2009 Mar 27.

Bystander effect-induced mutagenicity in HPRT locus of CHO cells following BNCT neutron irradiation: characteristics of point mutations by sequence analysis.

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Research Reactor Institute, Kyoto University, Kumatori-cho, Sennan-gun, Osaka, Japan.


To investigate bystander mutagenic effects induced by alpha particles during boron neutron capture therapy (BNCT), we mixed cells that were electroporated with borocaptate sodium (BSH), which led to the accumulation of (10)B inside the cells, with cells that did not contain the boron compound. BSH-containing cells were irradiated with alpha particles produced by the (10)B(n,alpha)(7)Li reaction, whereas cells without boron were only affected by the (1)H(n,gamma)(2)H and (14)N(n,rho)(14)C reactions. The frequency of mutations induced in the hypoxanthine-guanine phosphoribosyltransferase (HPRT) locus was examined in Chinese hamster ovary (CHO) cells irradiated with neutrons (Kyoto University Research Reactor: 5 MW). Neutron irradiation of 1:1 mixtures of cells with and without BSH resulted in a survival fraction of 0.1, and the cells that did not contain BSH made up 99.4% of the surviving cell population. Using multiplex polymerase chain reactions (PCRs), molecular structural analysis indicated that most of the mutations induced by the bystander effect were point mutations and that the frequencies of total and partial deletions induced by the bystander effect were lower than those resulting from the alpha particles produced by the (10)B(n,alpha)(7)Li reaction or the neutron beam from the (1)H(n,gamma)(2)H and (14)N(n,rho)(14)C reactions. The types of point mutations induced by the BNCT bystander effect were analyzed by cloning and sequencing methods. These mutations were comprised of 65.5% base substitutions, 27.5% deletions, and 7.0% insertions. Sequence analysis of base substitutions showed that transversions and transitions occurred in 64.7% and 35.3% of cases, respectively. G:C-->T:A transversion induced by 8-oxo-guanine in DNA occurred in 5.9% of base substitution mutants in the BNCT bystander group. The characteristic mutations seen in this group, induced by BNCT alpha particles, differed from those typical of gamma ray radiation.

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