Impact of the p53 status of the tumor cells on the effect of reactor neutron beam irradiation, with emphasis on the response of intratumor quiescent cells.
Human head and neck squamous cell carcinoma cells transfected with mutant p53 (SAS/mp53) or with neo vector as a control (SAS/neo) were inoculated subcutaneously into both the hind legs of Balb/cA nude mice. Tumor-bearing mice received 5-bromo-2'-deoxyuridine (BrdU) continuously to label all proliferating (P) cells in the tumors. After administration of sodium borocaptate-10B (BSH) or p-boronophenylalanine-10B (BPA), the tumors were irradiated with neutron beams. The tumors not treated with 10B-compound were irradiated with neutron beams or gamma-rays. The tumors were then excised, minced and trypsinized. The tumor cell suspensions thus obtained were incubated with a cytokinesis blocker, and the micronucleus (MN) frequency in cells without BrdU labeling (=quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. Meanwhile, 6 h after irradiation, tumor cell suspensions obtained in the same manner were used for determining the frequency of apoptosis in Q cells. The MN and apoptosis frequencies in total (P+Q) tumor cells were determined from the tumors that were not pretreated with BrdU. Without 10B-carriers, in both tumors, the relative biological effectiveness of neutrons was greater in Q cells than in total cells, and larger for low than high cadmium ratio neutrons. With 10B-carriers, the sensitivity was increased for each cell population, especially for total cells. BPA increased both frequencies for total cells more than BSH. Nevertheless, the sensitivity of Q cells treated with BPA was lower than that of BSH-treated Q cells. These sensitization patterns in combination with 10B-carriers were clearer in SAS/neo than in SAS/mp53 tumors. The p53 status of the tumor cells had the potential to affect the response to reactor neutron beam irradiation following 10B-carrier administration.
Masunaga
Shin-ichiro
S
Radiation Oncology Research Laboratory, Research Reactor Institute, Kyoto University, Sennan-gun, Osaka 590-0494, Japan. smasuna@rri.kyoto-u.ac.jp
Ono
Koji
K
Takahashi
Akihisa
A
Sakurai
Yoshinori
Y
Ohnishi
Ken
K
Kobayashi
Toru
T
Kinashi
Yuko
Y
Takagaki
Masao
M
Ohnishi
Takeo
T
D000818:Animals; D017209:Apoptosis / Q000528:radiation effects; D001894:Borohydrides / Q000627:therapeutic use*; D001896:Boron Compounds / Q000627:therapeutic use*; D016754:Boron Neutron Capture Therapy; D002294:Carcinoma, Squamous Cell / Q000473:pathology / Q000532:radiotherapy*; D002470:Cell Survival / Q000528:radiation effects; D005260:Female; D016158:Genes, p53 / Q000502:physiology*; D006258:Head and Neck Neoplasms / Q000473:pathology / Q000532:radiotherapy*; D006801:Humans; D051379:Mice; D015162:Micronucleus Tests; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000627:therapeutic use*; D013438:Sulfhydryl Compounds / Q000627:therapeutic use*; D014407:Tumor Cells, Cultured
C436045:4-dihydroxyborylphenylalanine; D001894:Borohydrides; D001896:Boron Compounds; D013438:Sulfhydryl Compounds; C014651:mercaptoundecahydrododecaborate; D010649:Phenylalanine