Effects of c-Myc overexpression on the DNA damage-induced G2/M checkpoint were studied in finite lifespan, normal human mammary epithelial cells (HMECs). Previously, we showed that c-Myc attenuates ...G1/S arrest and leads to an inappropriate entry of cells with damaged DNA into the S phase, following treatment with ionising radiation (IR). Here we show that, in striking contrast to control cells, c-Myc-overexpressing HMECs demonstrate a significant attenuation of the G2/M arrest, following IR, and enter into inappropriate mitoses. At the molecular level, ectopic overexpression of c-Myc leads to an unusually high level of expression of cyclin B1, and the elevated levels of cyclin B1 were maintained, after gamma-irradiation. Introduction of DNA damage in c-Myc-overexpressing, normal mammary epithelial cells eventually induces apoptosis, indicating a dramatic sensitisation by c-Myc of DNA damage-induced apoptosis. These two remarkable phenotypes, checkpoint attenuation and sensitisation to apoptosis, resulting from a deregulation of the protooncogene c-myc, may produce a unique pattern of alternating cycles, consisting first of amplification of DNA damage, followed by apoptosis-assisted selective pressure. The result of this alternating pattern of damage apoptosis could facilitate the selection of certain genomic alterations required for cellular survival and cellular transformation.
There is an increasing interest in the use of inhomogeneity corrections for lung, air, and bone in radiotherapy treatment planning. Traditionally, corrections based on physical density have been ...used. Modern algorithms use the electron density derived from CT images. Small fields are used in both conformal radiotherapy and IMRT, however, their beam characteristics in inhomogeneous media have not been extensively studied. This work compares traditional and modern treatment planning algorithms to Monte Carlo simulations in and near low-density inhomogeneities. Field sizes ranging from
0.5
cm
to
5
cm
in diameter are projected onto a phantom containing inhomogeneities and depth dose curves are compared. Comparisons of the Dose Perturbation Factors (DPF) are presented as functions of density and field size. Dose Correction Factors (DCF), which scale the algorithms to the Monte Carlo data, are compared for each algorithm. Physical scaling algorithms such as Batho and Equivalent Pathlength (EPL) predict an increase in dose for small fields passing through lung tissue, where Monte Carlo simulations show a sharp dose drop. The physical model-based collapsed cone convolution (CCC) algorithm correctly predicts the dose drop, but does not accurately predict the magnitude. Because the model-based algorithms do not correctly account for the change in backscatter, the dose drop predicted by CCC occurs farther downstream compared to that predicted by the Monte Carlo simulations. Beyond the tissue inhomogeneity all of the algorithms studied predict dose distributions in close agreement with Monte Carlo simulations. Dose–volume relationships are important in understanding the effects of radiation to the lung. The dose within the lung is affected by a complex function of beam energy, lung tissue density, and field size. Dose algorithms vary in their abilities to correctly predict the dose to the lung tissue. A thorough analysis of the effects of density, and field size on dose to the lung and how modern dose calculation algorithms compare to Monte Carlo data is presented in this research project. This work can be used as a basis to further refine an algorithm’s accuracy in low-density media or to correct prior dosimetric results.
It has been estimated that approximately 1% of the general population are ataxia telangiectasia (AT) mutated (ATM) heterozygotes. The ATM protein plays a central role in DNA-damage response pathways; ...however, the functional consequences of the presence of either heterozygous truncating or missense mutations on ATM expression and the ionising radiation (IR)-induced cellular phenotype remain to be fully determined. To investigate this relationship, the ATM mRNA and protein levels and several cellular end points were characterised in 14 AT heterozygote (AT het) lymphoblastoid cell lines, compared to normal and AT homozygote lines. The AT het cell lines displayed a wide range of IR-induced responses: despite lower average levels of ATM mRNA and protein expression compared to normal cells, 13 out of 14 were capable of phosphorylating the ATM substrates p53-ser15 and Chk2, leading to a normal cell cycle progression after irradiation. However, cell survival was lower than in the normal cell lines. The presence of a missense compared to a truncating mutation was associated with lower cell survival after exposure to 2 Gy irradiation (P=0.005), and a higher level of ATM mRNA expression (P=0.047). Our results underline the difficulty in establishing a reliable test for determining ATM heterozygosity.
The aim of the study was to compare cross sections with longitudinal sections in histopathological examination of the rat heart after irradiation, to find the most optimal method for the detection of ...cardiac radiation injuries. For this purpose, rats were irradiated locally on the heart with a single dose of 0 or 20 Gy. At different time points after irradiation, hearts were perfused and cut into longitudinal or cross sections. In both sections, several histopathological changes were scored on a graded scale between 0 and 3. Mast cells, which are thought to play a role in tissue remodelling, were counted. After 20 Gy, frequently occurring lesions were most severe in the upper half of the ventricles and the septum. These lesions could only be detected when using longitudinal sections, resulting in a higher total histopathological score than the examination of a single cross section. From 3 months onwards, changes in coronary arteries of irradiated hearts included endothelial cell loss, a loss of smooth muscle cells and fibrosis in media and adventitia. Up to 1 month after irradiation, mast cell densities of the left and right ventricles were decreased after 15 and 20 Gy, compared to time-matched controls, followed by increases from 3 months onwards. In the left ventricle, mast cell densities correlated with myocardial degeneration and fibre loss. The results of this study show that the usage of a single longitudinal section in the histopathological examination of the irradiated rat heart leads to the recognition of more severe injuries, including myocardial degeneration and fibrosis, in ventricular tissue than the usage of a single midventricular cross section. Morphological changes observed in coronary arteries of irradiated hearts might lead to a decreased compliance of the coronary artery wall. Further investigation is needed to determine the role of mast cells in cardiac tissue remodelling after irradiation.
A model based on the linear quadratic model that has been corrected for repopulation, sublethal cell damage repair, and RBE effect has been used to determine the prescription dose for prostate ...permanent brachytherapy using seeds loaded with a mixture of
Pd
103
and
I
125
or a mixture of
Pd
103
and
Cs
131
. The prescription dose was determined by comparing the tumor cell survival fractions between the considered biradionuclide seed implant and one monoradionuclide seed implant chosen from
Pd
103
,
I
125
, and
Cs
131
. Prostate edema is included in the model. The influence of the value of the radiobiological parameters and RBE were also investigated. Two mixtures of radionuclides were considered:
Pd
0.75
103
–
I
0.25
125
and
Pd
0.25
103
–
Cs
0.75
131
, where the subscripts indicate the fractions of total initial internal activity in the biradionuclide seed. These fractions were selected in order to obtain a dose distribution that lies between that of
Pd
103
and
I
125
∕
Cs
131
. As expected, the computed prescription dose values are dependent on the model parameters (edema half-life and magnitude, radiobiogical parameters, and RBE). The radionuclide used as a benchmark also has a strong impact on the derived prescribed dose. The large uncertainties in the radiobiological parameters and RBE values produce big errors in the computed prescribed dose. Averaged over the range of all the parameters and depending on the radionuclide used as a benchmark (in subscript), the derived prescription dose for the first mixture (PdI) would be:
D
Pd
Pd
I
=
142
−
16
+
15
Gy
and
D
I
Pd
I
=
142
−
8
+
6
Gy
; and
D
Pd
Pd
Cs
=
128
−
13
+
13
Gy
and
D
Cs
Pd
Cs
=
115
−
7
+
6
Gy
for the PdCs mixture. The uncertainties could be reduced if the radiobiological parameters and RBE value were known more accurately. However, as edema characteristics are patient dependent and can be obtained only after the treatment, an unpredictable error is unavoidable.
Ionising radiation-induced reactive oxygen species (ROS) overproduction induces keratinocyte alterations and constitutes one of the most common effects after therapeutic γ-irradiation. ROS production ...is controlled by a complex enzymatic system.
The aim of our study is to analyse the role of radiation-induced oxidative stress in keratinocytes death by apoptosis. We hypothesized that keratinocyte capacity to hamper radiation-induced ROS generation may control their radiosensitivity.
For this purpose, an original human skin explant model was developed and two types of human epidermal cells were used: primary keratinocytes NHEK and spontaneous non-tumourigenic cell line HaCaT.
cDNA-arrays analysis was performed 24
h after a 20
Gy γ-radiation and revealed down-regulation of genes involved in oxidative stress control and the apoptosis process. This was confirmed by alterations in catalase, GPx and SOD enzymatic activities. This redox modulation was concomitant to the down-regulation of anti-apoptotic genes and up-regulation of some pro-apoptotic genes (caspase 10, ubiquitin C). Interestingly TUNEL labelling revealed an increase in the number of apoptotic cells. We also demonstrated a differential inducibility of the cell antioxidant network in two keratinocyte lines, which results in a differential cellular level of ROS, explaining their different radiosensitivities.
Keratinocytes apoptosis is partly dependent on ROS production after exposure to γ-rays. In addition, the differential radiosensitivity of keratinocytes is linked to different oxidative stress responses.
For a retrospective dose estimation of human exposure to ionising radiation, a partial genome analysis is routinely used to quantify radiation-induced chromosome aberrations. For this purpose, ...fluorescence in situ hybridisation (FISH) with whole chromosome painting probes for selected chromosomes is usually applied covering about 20% of the whole genome. Since genome-wide screening techniques like spectral karyotyping (SKY) and multiplex FISH (mFISH) have been developed the detection of radiation-induced aberrations within the whole genome has now become feasible. To determine the correspondence between partial and whole genome analysis of radiation-induced chromosome aberrations, they were measured comprehensively in this study using in vitro irradiated blood samples from three donors. We were able to demonstrate that comparable results can be detected with both approaches. However, complex aberrations might be misinterpreted by partial genome analysis. We therefore conclude that whole genome analysis by SKY is useful especially in the high dose range to correct aberration data for complex exchange aberrations.
Ionising radiation is used for the treatment of pituitary tumours as fractionated radiotherapy, where the total dose reaching the tumour area is in the range of 40-50 Gy, or during stereotactic ...radiosurgery, where the total dose reaching the tumour area during one session is in the range of 20-90 Gy. In this study, we investigated the effect of ionising radiation of 60Co (dose rate of 3 Gy/min, similar to that used during gamma knife procedure) on the mode of cell death of the somatomammotroph pituitary cell line, GH3, an immortalized cell line derived from a rat pituitary adenoma. We found that the basic mechanism of cell death induced by irradiation of this GH3 cell line by γ-rays was programmed cell death-apoptosis. Doses of 20-50 Gy were shown to inhibit proliferation in these cells. 24 hours after irradiation with a dose of 20 and 50 Gy, cells were shown to accumulate in the G2/M phase of cell cycle. This cell cycle arrest lasted for at least ten days. Apoptosis was detected 72 hours towards until the end of the study (10 days). However, a significant number of cells were still alive ten days following irradiation. We conclude that ionising radiation doses of 20 and 50 Gy induce pituitary GH3 cell apoptosis following cell cycle arrest in the G2/M phase.
Cyclopentenyl cytosine (CPEC), targetting the de novo biosynthesis of cytidine triphosphate (CTP), increases the cytotoxicity of gemcitabine (2',2'-difluoro-2'-deoxycytidine, dFdC) alone and in ...combination with irradiation in several human tumour cells in vitro. We investigated whether CPEC enhances the therapeutic ratio of gemcitabine and irradiation in human pancreatic BxPC-3 xenografts and in rat syngeneic L44 lung tumours. These models were selected because gemcitabine and radiation are used to treat both pancreatic and lung cancer patients and both models differ in growth capacity and in gemcitabine-induced radiosensitisation. A profound dose-dependent CTP-depletion was observed after a single injection of CPEC in both tumour tissue and in normal jejunum. In both models, CPEC alone induced a slight but significant tumour growth delay. The combination of CPEC with gemcitabine, at time intervals that showed CTP-depletion after CPEC, enhanced neither tumour growth delay nor toxicity as compared to gemcitabine alone. In addition, no beneficial effect of CPEC was observed in combination with gemcitabine and radiation. These results suggest that CPEC and gemcitabine alone as well as in combination with radiation target a similar cell population in both tumour models. In conclusion, future clinical development of CPEC as a modulator of gemcitabine combined with radiation is unlikely.
Poly(siloxaneurethaneureas) (PSURURs) prepared from aromatic and aliphatic isocyanates were investigated upon exposure to ionising radiation. Radicals are formed both in siloxane and urethane ...segments. In comparison with aliphatic analogues it was found that in aromatic PSURURs: (1) concentration of all radicals is lower, (2) relative concentration of methylene radicals formed in siloxane units is higher, (3) the radiation yield of H
2 is more than three times smaller and (4) it seems that efficiency of cross-linking is less significant.
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