The two radiosensitizers SR-2508 (etanidazole) and paclitaxel (taxol) have different dose-limiting toxicities in humans. Combination of the two radiosensitizers may increase radiosensitization without increasing toxicity. This study was carried out to determine the synergistic radiosensitizing effect of combination of SR-2508 and paclitaxel in two hypoxic human tumor cell lines: a breast carcinoma (MCF-7) and a carcinoma cervicis (HeLa). The 3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay was used to determine the number of surviving cells. Cell cycle was evaluated by flow cytometry. Cell viability was measured by the ability of single cells to form colonies in vitro. Our data demonstrated that the radiosensitization produced by the two radiosensitizers was additive in hypoxic HeLa cells while held in the G1 phase of the cell cycle. On the other hand, there was no synergistic radiosensitizing effect in hypoxic MCF-7 cells by combination of the two drugs. Our results suggested that the synergistic radiosensitizing effect of SR-2508 and paclitaxel may be tumor-dependent and that breast cancer may not be a good candidate. This study may provide a new combination of radiosensitizers in radiotherapy for cervical carcinoma.
The objective of this study was to explore whether a specific inhibitor of PI3K, wortmannin, could potentiate the antitumor effect of radiation in vivo, particularly on radioresistant murine tumors. C3H/HeJ mice bearing syngeneic hepatocarcinoma (HCa-I) were treated with 25 Gy radiation, wortmannin, or both. Wortmannin was administered intraperitoneally (1 mg/kg) once daily for 14 days. Tumor response to treatment was determined by a tumor growth delay assay. Possible mechanisms of action were explored by examining the level of apoptosis and regulating molecules. The expression of regulating molecules was analyzed by Western blot for p53 and p21WAF1/CIP1, and immunohistochemical staining for p21WAF1/CIP1, CD31 and VEGF. In the tumor growth delay assay, wortmannin increased the effect of tumor radioresponse with an enhancement factor (EF) of 2.00. The level of apoptosis achieved by the combined treatments was shown to be no more than an additive effect; peak apoptotic index was 11% in radiation alone, 13% in wortmannin alone, and 19% in the combination group. Markedly increased areas of necrosis at 24 h in the combination group were noted. Western blotting showed upregulation of p21WAF1/CIP1 in the combination treatment group, which correlated with low levels of VEGF. Microvascular density was evidently also reduced, based on low expression of CD31. In murine hepatocarcinoma, the antitumor effect of radiation was potentiated by wortmannin. The mechanism seems to involve not only the increase of induced apoptosis but also enhanced vascular injury. Wortmannin, in combination with radiation therapy, may have potential benefits in cancer treatment.
This study evaluated micronuclei induction in peripheral T-lymphocytes after BNCT (boron neutron capture therapy) irradiation of 15 brain tumor and 20 head-and-neck cancer patients. In all of these patients, the micronucleus frequency increased after BNCT; the number of micronuclei per 1000 binucleated T-lymphocytes after BNCT increased by 33.0 ± 12.2 for brain tumor patients and 22.8 ± 10.3 for head-and-neck cancer patients. In 14 thyroid cancer patients who were administered radioiodine (3.3-5.6 GBq) treatment, the frequency of micronuclei after internal radioiodine therapy increased to 105.0 ± 30.5 per 1000 binucleated T-lymphocytes. The increased micronucleus frequency of BNCT patients was less than one-third that seen for thyroid cancer patients after radioiodine treatment. These results demonstrate the usefulness of BNCT for selective high-LET radiotherapy, in association with a low irradiation effect of cytological radiation damage after BNCT, which provides a high tumor target dose for each cancer patient.
This study was undertaken to investigate induction of tumors by monoenergetic neutrons in B6C3F1 mice. Individual groups of 6 week-old animals of both sexes (about 30 mice/group) were exposed to 0.5 Gy of various monoenergetic neutrons (dose rate 0.5 cGy/min) and then observed for 13 months. The incidences of tumors (mainly liver neoplasms) in non-irradiated male and female controls were 11% and 0%, respectively. In the irradiated animals, the incidences were 53%, 50%, 60% and 43% in males, and 75%, 81%, 71%, and 85% in females, after 0.18, 0.32, 0.6 and 1.0 MeV neutron exposure, respectively. There were no significant differences in the tumor induction rate among the different energy groups.
Nasopharyngeal carcinoma (NPC) is one of the common malignant tumors in China. Radiotherapy and chemotherapy are the main therapy methods for NPC. To enhance the specific antitumor effect, a novel vector with radiosensitivity and tumor specificity was constructed in this study, which enables the reduction of dosage of radiation and chemotherapeutic drugs due to its double killing effect. Four DNA elements, Egr-1 promoter, Cytosine deaminase (CD) gene, hTERT promoter, Survivin antisense oligonucleotides were amplified and constructed in pcDNA3.1 vector. CD and Survivin gene expression in CNE-2 cells were detected by RT-PCR. High performance liquid chromatography (HPLC) was employed to determine the transformation from the prodrugs 5-FC to 5-FU. Hoechst33258 staining of the nuclei and methylthiazolyl tetrazolium(MTT) assay were applied to detect apoptosis and cell survivability, respectively. In addition, the anti-tumor effects were examined in vivo by injecting cells with different vectors into nude mice. Our results revealed a notable killing effect of combined treatment with 5-FC and radiation on CNE-2 cells transfected with vectors in vitro. This effect was especially notable on pEC-TS transferred cells, which showed 57% of cells were killed. In vivo, an obvious suppression of tumor was displayed in pEC-TS group, which was significantly different from other groups (p < 0.05). Consequently, this expression cassette may have a great therapeutic potential for the treatment of NPC.
Werner syndrome (WS) is a premature aging syndrome caused by mutations of the WRN gene. Here, we demonstrate that a strain of WS fibroblast cells shows abnormal karyotypes characterized by several complex translocations and 50-fold more frequency of abnormal metaphases including dicentric chromosomes without fragments than normal cells when examined at a similar culture stage. Further, telomere fluorescence in situ hybridization indicates that the abnormal signals, extra telomere signal and loss of telomere signal, emerge two- to three-fold more frequently in WS cells than in normal cells. Taken together, these results indicate that chromosome instability including dysfunction of telomere maintenance is more prominent in WS cells than in normal cells. In addition, the accumulation of DNA double-strand breaks (DSBs) at the G1 phase, including those at telomeres, detected by phosphorylated ATM (ataxia telangiectasia mutated) foci is accelerated in WS cells even at a low senescence level. The increased accumulation of DSBs in WS cells is reduced in the presence of anti-oxidative agents, suggesting that enhanced oxidative stress in WS cells is involved in accelerated accumulation of DSBs. These results indicate that WS cells are prone to accumulate DSBs spontaneously due to a defect of WRN, which leads to increased chromosome instability that could activate checkpoints, resulting in accelerated senescence.
Until now, the multiple biological effects of ionizing radiation on liver have been reported. However, there has not been any reports of fast neutron-mediated liver injuries including liver regeneration or fibrosis. Here, we described the biological effects of acute fast neutron irradiation on the liver. After the fast neutron irradiation of 0, 0.25, 1, 2, 4 and 8 Gy on mice, hepatocyte necrosis and a decrease in the total number of hepatocytes were induced dose-dependently. Binucleated hepatocytes and PCNA positive hepatocytes increased significantly at 0.25 and 1 Gy, but decreased markedly at 2, 4 and 8 Gy. The expression of cytochrome P450 2E1 (CYP2E1) showed a dose-dependent increase after fast neutron irradiation. The activation of p-Smad2/3, signaling intermediates of transforming growth factor-beta (TGF-β), increased in hepatocytes after exposure of 0.25, 1, and 2 Gy of fast neutrons, but it was not detected in hepatic stellate cells (HSCs). In conclusion, fast neutron-induced liver damages, likely loss of hepatocytes, necrotic foci and vacuolar changes, were note on the dose dependent manner and hepatocellular regeneration were significantly diminished at doses of 2, 4 and 8Gy in a dose-dependent manner. These alterations may at least in part be associated with dose-dependent increase in CYP2E1 and p-Smad2/3. These results show promise as an approach for the treatment of fast neutrons on liver tumors and in the study of pathogenesis regarding the fast neutron-irradiated damages of the liver.
The effect of mononuclear copper (II) complex of curcumin in 1:1 stoichiometry (hereafter referred to as complex) administered 30 min before γ-irradiation (4.5 Gy) on alterations in antioxidant and Thiobarbituric acid reactive substances (TBARS) levels in livers was studied in comparison to curcumin at a dose of 50 mg/kg. The different antioxidants like GSH, GST, catalase, SOD, TBARS and total thiols were estimated in the liver homogenates excised at different time intervals (1, 2 and 4 h) post irradiation using colorimetric methods. There was a radiation-induced decrease in the levels of all the studied enzymes at 1 h post irradiation, while an increase was observed at later time points. Both curcumin and complex treatment in sham-irradiated mice decreased the levels of GSH and total thiols, whereas there was an increase in the levels of catalase, GST and SOD compared to normal control. Under the influence of irradiation, both curcumin and complex treatment protected the decline in the levels of GSH, GST, SOD, catalase and total thiols, and inhibited radiation-induced lipid peroxidation. Further, the complex was found to be more effective in protecting the enzymes at 1 h post irradiation compared to curcumin treated group. This may be due to the higher rate constants of the complex compared to curcumin for their reactions with various free radicals.
To carry out the radio-microsurgery study using silkworm, Bombyx mori, we have already developed the specific irradiation systems for eggs and third to fifth instar larvae. In this study, a modified application consisting of the first instar silkworm larvae was further developed using heavy-ion microbeams. This system includes aluminum plates with holes specially designed to fix the first instar silkworm larvae during irradiation, and Mylar films were used to adjust energy deposited for planning radiation doses at certain depth. Using this system, the suppression of abnormal proliferation of epidermal cells in the knob mutant was examined. Following target irradiation of the knob-forming region at the first instar stage with 180-μm-diameter microbeam of 220 MeV carbon (12C) ions, larvae were reared to evaluate the effects of irradiation. The results indicated that the knob formation at the irradiated segment was specially suppressed in 5.9, 56.4, 66.7 and 73.6% of larvae irradiated with 120, 250, 400 and 600 Gy, respectively, but the other knob formations at the non-irradiated segments were not suppressed in either irradiation. Although some larva did not survive undesired non-targeted exposure, our present results indicate that this method would be useful to investigate the irradiation effect on a long developmental period of time. Moreover, our system could also be applied to other species by targeting tissues, or organs during development and metamorphosis in insect and animals.
The geometric locations of ion traversals in mammalian cells constitute important information in the study of heavy ion-induced biological effects. We employed a contact microscopy technique, which was developed for boron imaging in boron neutron capture therapy to the irradiation mammalian cells by low-energy heavy ions. This method enables the simultaneous visualization of mammalian cells as a relief on a plastic track detector, CR-39, and the etch pits which indicate the positions of ion traversals. This technique provides visual geometric information about the cells and ion traversal, without any specially designed devices or microscopes. Only common laboratory equipment, such as a conventional optical microscope, a UV lamp, and commercially available CR-39 is required. To validate this method, CHO-K1 and HeLa cells were cultured on the CR-39 surface and then irradiated with low-energy Ar and Ne ions, respectively. The positions of induced DNA double strand breaks were detected as γ-H2AX fluorescent spots, which coincided with the positions of the etch pits in the cell relief image.