Оценка последствий дистанционной лучевой терапии у больных раком лёгкого путём анализа хромосомных аберраций в лимфоцитах крови Хвостунов И.К. 1 , Курсова Л.В. 1 , Севанькаев А.В. 1 , Рагулин Ю.А. 1 , Шепель Н.Н. 1 , Коровчук О.Н. 1 , Пятенко В.С. 1,2, Хвостунова Т

The incidence of unstable chromosome aberrations in peripheral blood lymphocytes from unirradiated control subjects was analyzed using cytogenetic data obtained from 9 cytogenetic laboratories located in Moscow, St.-Petersburg, Obninsk, and Dubna (Russia). The objective of this study was to estimate the level and spectrum of spontaneous chromosome aberrations in human lymphocytes. 1140 blood samples were taken from 1112 subjects (594 men and 546 women) aged 1 to 72. The total metaphase number was 466795. The uniform Giemsa method for peripheral blood lymphocyte cultures was used. After counting 466795 metaphases, 4288 chromosomal aberrations of various types were classified. The most frequent types of aberrations were acentrics and chromatid deletions. They made up 90% of the total number of aberrations. The remaining 10% were exchange aberrations. The number of chromosome exchanges (dicentrics and centric rings) was twice the number of chromatid exchanges. Overall, the portion ofcells with chromosomal or (and) chromatid aberrations was 0.89 +/- 0.01%; the frequency of acentrics was 0.29 +/- 0.01; the frequency of dicentrics was 0.046 +/- 0.003; the frequency of unstable chromosome aberrations was 0.35 +/- 0.01; and the frequency of chromatid aberrations was 0.57 +/- 0.01 per 100 cells.

The incidence of unstable chromosome aberrations in peripheral blood lymphocytes from unirradiated control subjects was analyzed using cytogenetic data obtained from 9 cytogenetic laboratories located in Moscow, St.-Petersburg, Obninsk, and Dubna (Russia). The objective of this study was to estimate the level and spectrum of spontaneous chromosome aberrations in human lymphocytes. 1140 blood samples were taken from 1112 subjects (594 men and 546 women) aged 1 to 72. The total metaphase number was 466795. The uniform Giemsa method for peripheral blood lymphocyte cultures was used. After counting 466795 metaphases, 4288 chromosomal aberrations of various types were classified. The most frequent types of aberrations were acentrics and chromatid deletions. They made up 90% of the total number of aberrations. The remaining 10% were exchange aberrations. The number of chromosome exchanges (dicentrics and centric rings) was twice the number of chromatid exchanges. Overall, the portion ofcells with chromosomal or (and) chromatid aberrations was 0.89 +/- 0.01%; the frequency of acentrics was 0.29 +/- 0.01; the frequency of dicentrics was 0.046 +/- 0.003; the frequency of unstable chromosome aberrations was 0.35 +/- 0.01; and the frequency of chromatid aberrations was 0.57 +/- 0.01 per 100 cells.

The yield of chromosome aberrations induced by gamma-radiation of 60Co in human blood lymphocytes in vitro at low doses (30 divided by 600 mGy) and low dose rates (0.70, 5.05, 59.2 mGy/min) was investigated. It was found that the observed level of chromosomal aberrations induced by gamma-irradiation was unaffected by the value of the dose rate when using constant dose rate and obtaining different doses by altering the exposure time. However, a relatively enhanced level of chromatid aberrations was found at 5.05 and 59.2 mGy/min dose rates in the dose range less than 250 mGy. We have found that the observed level of the sum of chromosomal aberrations induced by gamma-irradiation at doses less than 250 mGy and a dose rate of 59.2 mGy/min was essentially larger compared with the level extrapolated from high doses (above 300 mGy) using a linear-quadratic dose curve. This complied with our previous finding in 1976, 1977 when the enhanced level of dicentrics was only found at a high dose rate approximately 500 mGy/min. Such a non-linear cytogenetic effect does not manifest itself statistically significantly at dose rates of 0.70 and 5.05 mGy/min for the sum of chromosomal aberrations and does not manifest itself at all for dicentrics at all the examined dose rates.

The objective of this study was to investigate in vivo the dose response of radiation induced chromosomal aberrations in human blood lymphocytes of lung cancer patients given non-uniform fractional exposures to high doses of therapeutic 60Co gamma-rays delivered synchronously with polychemotherapy. The chromosome aberration analysis was carried out in peripheral blood lymphocytes of 13 lung cancer patients who manifested II to IV developmental clinical stage. During the course of radiotherapy they received the accumulated tumor dose ranged 47.5 to 70 Gy. The yield ofdicentrics, centric rings and fragments was measured in the blood samples taken before treatment, after the first day and after the complete course of radiotherapy. Based on cytogenetic measurements of 3 patients, the average tumor dose after the first day was estimated to be 2.1 to 3.0 Gy given that the corresponding physical dose was (1.0 Gy + 1.5 Gy). The quotient of the individual dose estimated by the frequency of aberrations to the physical dose after the complete course of radiotherapy was calculated for all 13 patients. The mean quotient was shown to be equal to 93 +/- 9% ranged 50 to 154%.

мГр/мин. Для частоты общего числа аберраций при мощностях дозы 0.70 и 5.05 мГр/мин, а также для дицентриков при всех мощностях дозы нелинейный эффект на дозовой зависимости практически отсутствует. льших доз. Обнаружено, что нелинейный эффект, выражающийся в наличии плато на дозовой кривой в диапазоне 30?250 мГр, явно проявляется только для общей частоты аберраций хромосомного типа при одной мощности дозы – 59.2 мГр/мин, в отличие от полученных ранее результатов [1, 2], где эффект плато проявлялся для частоты дицентриков при мощности дозы 500 мГр/мин. Для частоты общего числа аберраций при мощностях дозы 0.70 и 5.05 мГр/мин, а также для дицентриков при всех мощностях дозы нелинейный эффект на дозовой зависимости практически отсутствует.

Using routine methods over 25 years, changes in the registered levels of chromosomal aberrations were studied in the peripheral blood lymphocyte cultures of 74 patients who were irradiated as a result of the Chernobyl accident. The initial dosage estimations by mean dicentric frequency varied from 0.2 to 9.8 Gy. Generally, a double exponential type model was most adequate for the quantitative description of the elimination of cytogenetic indexes associated with different types of unstable chromosomal aberrations. Great individual variability of the elimination rate of chromosomal aberrations and its dependency on the value of the originally estimated dosage were found during the first period. A computer method for retrospective dos-age estimation was developed based on this data. The method is based on analysis of cell distributions in accordance with the number of dicentrics and, as a whole, unstable chromosomal aberrations contained in them. In addition, the dynamics of the translocation frequencies in the peripheral blood lymphocyte cultures of a number of patients from this contingent were investigated, beginning at 10 years after the irradiation using the FISH method of chromosome staining.

Changes of registered levels of chromosome aberrations were studied in the peripheral blood lymphocyte cultures of 74 patients irradiated as a result of the Chernobyl accident by the instrumentality of the routine method during 25 year. The initial dose estimations by average dicentrics frequency varied from 0.2 to 9.8 Gy On the whole, the model of a double exponential type was the most adequate for the quantitative description of elimination of cytogenetical indices associated with different types of unstable chromosome aberrations. High individual variability of the elimination rate of chromosome aberrations and its dependency from the value of originally evaluated dose were discovered in the first period. The computer method of retrospective dose estimation was developed on the basis of this material. The method is based on the analysis of cell distributions in accordance with the number of dicentrics and as a whole, unstable chromosome aberrations, contained in them. In addition the dynamics of translocation frequencies in the peripheral blood lymphocyte cultures of a number of patients from this contingent was investigated starting from 10 years after irradiation by the instrumentality of FISH-method of chromosome staining.

This paper considers how well standard calibration curve for translocations constructed for lymphocyte cultures irradiated in vitro with gamma-rays from (60)Co compares with the translocations yield in lymphocytes taken from people at a long post-exposure time. Data were used from radiation accident victims overexposed to doses ranging from 0.2 to 8.5 Gy and who were cytogenetically followed-up for various times upto 50 y. Their cultured lymphocytes had been scored both by the conventional dicentric method and by FISH for all translocations involving painted chromosomes (2, 3, 8); (2, 3, 5) or (2, 4, 12). The in vivo dose response relationship was derived by fitting translocation frequencies to the contemporary individual doses obtained independently and confirmed by different biological assays and physical dosimetry. A comparison with the conventional in vitro curve indicates reductions of translocation frequencies with increasing time which would prejudice retrospective dose assessment by FISH. This has led to the possibility to amend the in vitro dose response curve for translocations to make it more suitable for use in retrospective biodosimetry. This approach for retrospective biodosimetry therefore uses a dose response relationship based on truly persisting translocations. (C) 2011 Elsevier Ltd. All rights reserved.

The study was based on the examination of tumors in 31 patients radically operated on for locally advanced gastric cancer at the Medical Radiology Research Center, Russian Academy of Medical Sciences, in 2000-2007. The pilot study indicated rather high rates of Her-2/neu hyperexpression and amplification. It was shown that immunohistochemical and FISH assays used in breast cancer might be successfully used in gastric cancer. Analysis demonstrated that Her-2/neu hyperexpression/amplification in patients with gastric cancer was an important factor of poor prognosis. The authors suggested for the first time that Her-2/neu might be considered as one of the determinants of the mechanisms of metastasis in patients with gastric cancer.

Tumor markers for diagnosis of bladder cancer and defining the prognosis of the disease have been actively studied for past decade. FISH-study demonstrates the highest sensitivity and specificity. Abbot Company has developed a diagnostic test system ≪UroVision≫ which includes colored DNA-probes referring to 3, 7, 17 and 9p21 chromosomes. This system use provides clinicians with insights into an early diagnosis, prognosis and monitoring of efficacy of anticancer therapy

Patients with primary diagnosis of bladder cancer (42 persons) were studied. FISH analysis performed in all patients showed the following results: 39 (93%) patients were FISH positive and 3 (7%) were FISH negative. The molecular cytogenetic criterion for determining the grade of differentiation of Ta and T1 tumors chromosomal hyperploidy in morphologically abnormal cells, was suggested. All patients demonstrating increased amount (more than 26%) of abnormal cell with hyperploidy of chromosomes 3, 7 and 17 were found to have high grade malignancies.

Summarized results of 5 repeated experiments of comparative study of radiation effects of the pulse reactor BARS-6 either in a single pulse or a continuos irradiation mode on human lymphocytes are presented. Higher efficiency (30-40%) of continuous irradiation (exposure duration 1 h) rather than pulse irradiation with ultra-high dose rates (1-2.5) x 10(6) Gy/min (pulse duration 65 micros) was confirmed. The efficiency ratio did not depend on the temperature, 20 degrees C or 0 degrees C, during the exposure. Cell repair system and chromatin conformation influence on the results obtained is discussed.

The results of comparative analysis of gene and structural mutations found in peripheral blood lymphocytes of inhabitants of Orel district areas contaminated with radionuclides as a result of Chernobyl accident are presented. The average level of 137Cs contamination in those areas ranged about 22-113 kBq/m2. In the study group was found the enhanced frequency of somatic cells with gene and structural mutations compared with laboratory control level by synchronous applying a T-cell receptor (TCR) loci mutation assay and cytogenetic analysis of unstable aberrations. The case-control comparison was carried out using the measured mutation frequencies and cases of various thyroid gland sickness recognized by ultrasonic examination. The cytogenetic assay did not show the statistical difference between healthy group and subjects with thyroid gland sickness. The average frequency of TCR loci mutation cells in the subjects with thyroid gland sickness was found to be statistically higher comparing with healthy persons. This finding was true for each study region and for Orel district in total. The subgroup of subject exposed in utero in 1986, soon after accident was analyzed. Both cytogenetic and TCR loci mutation assays shown enhancement of average mutation frequency in somatic cells in the subjects of this subgroup with thyroid gland sickness comparing with healthy persons.

The paper presents the results of examination of children and teenagers living on the contaminated areas of Oryol (248 subjects) and Kaluga (224 subjects) oblasts since the Chernobyl accident occurred. The goal of the study was carrying out a comparative analysis the results of cytogenetic assay and estimation of morphologic-function status of thyroid glands by ultrasonic exclusion. The study was consisted of synchronous ultrasonic diagnosis and cytogenetic analysis of chromosomal aberrations in peripheral blood lymphocytes performed in 1998 and 2003 years in two oblasts respectively. Based on combined cytogenetic data it was found no significant discrepancy between healthy and thyroid gland pathology subjects studied in both oblasts. At once the subgroup with high level of chromosomal aberrations exhibited elevated percentage of thyroid diseases compared with the subgroup with low level of chromosomal aberrations, namely by 15% for Kaluga and by 25 % for Oryol oblasts. The highest level of thyroid gland disease incidence was found in the subgroup of subjects with high level of chromosomal aberrations who was born in 1986-1987 years, that is by 44 % for Kaluga and by 110 % for Oryol oblasts.

Molecular-biochemical and cytogenetic analyses were made on blood cells of 17 radiation accident victims who, from 1.7 to 43.8 years previously, had suffered acute radiation sickness (ARS) ranging from severity grades I to IV. Molecular-biochemical data were obtained with patients' leukocytes and with mononuclear cells on their oxidative status by a) the level of an anion-radical O2*- in the 3-(4,5-dimethyl-2-thiazol-2-yl)-2,5-diphenyl-2-tetrazolium bromide (MTT) test and b) the sum of reactive oxygen species in the 2,7-dichlorodihydrofluorescein diacetate (DCFDA) test together with a test for DNA strand unwinding in alkaline conditions by measurements of fluorescence intensity of ethidium bromide. Each parameter was measured in freshly sampled cells and during a 5-hour incubation as absolute means on 5 measurements and in % change from the initial values. Cytogenetic data were obtained from the standard metaphase preparations scored for routine unstable chromosomal aberrations (us-CA)--dicentrics; and stable aberrations (sCA)--translocations. The latter ones were assayed by the FISH method using whole chromosome 2, 4 and 12 fluorescent probes and scaled up to genome equivalence. For all patients reduced oxidative status of about 25-30% was obtained by the MTT-test (p < 0.005), and the DCFDA-test (p < 0.027). The yield of usCA depended on the time after irradiation with higher yields associated with the shorter postirradiation times and reducing almost to expected background frequencies. The yield of sCA was high for all patients, correlating with the severity of ARS whilst the molecular-biochemical parameters showed no relationship with ARS. A correlation was observed between parameters of oxidative status and % of cells with usCA: by the MTT-test r = from 0.50 up to 0.61 (p = from 0.06 up to 0.003), but by the DCFDA-test the strength of correlation was smaller: r = from 0.38 up to 0.48. An inverse correlation was found between initial oxidative state of mononuclear cells and the frequency of CA in lymphocytes. Similarly a marked inverse correlation between degree of DNA unwinding by the ethidium bromide assay on leukocytes and sCA in lymphocytes was also noted. The feasibility of radiation-induced delayed genomic instability in vivo for humans long time after irradiation is discussed.

The results of the comparative study of radiation effects of the pulse reactor BARS-6 either in single pulse or continuos irradiation mode on human G0 lymphocytes are presented. Under identical doses the cytogenetic efficiency was observed to be higher for continuous irradiation (1 hour) than for single pulse irradiation with ultrahigh dose rate (0.5-3) x 106 Gy/minutes (pulse duration 65 x 10(-6) s). The difference averaged about 37% on total aberration frequency and 27% on the sum of dicentrics and centric rings. The influence of the dose rate and of the mixed gamma-neutron irradiation on the obtained results is discussed.