Showing papers on "Sister chromatid exchange published in 2015"

Defective sister chromatid cohesion is synthetically lethal with impaired APC/C function.

Abstract: Warsaw breakage syndrome (WABS) is caused by defective DDX11, a DNA helicase that is essential for chromatid cohesion. Here, a paired genome-wide siRNA screen in patient-derived cell lines reveals that WABS cells do not tolerate partial depletion of individual APC/C subunits or the spindle checkpoint inhibitor p31(comet). A combination of reduced cohesion and impaired APC/C function also leads to fatal mitotic arrest in diploid RPE1 cells. Moreover, WABS cell lines, and several cancer cell lines with cohesion defects, display a highly increased response to a new cell-permeable APC/C inhibitor, apcin, but not to the spindle poison paclitaxel. Synthetic lethality of APC/C inhibition and cohesion defects strictly depends on a functional mitotic spindle checkpoint as well as on intact microtubule pulling forces. This indicates that the underlying mechanism involves cohesion fatigue in response to mitotic delay, leading to spindle checkpoint re-activation and lethal mitotic arrest. Our results point to APC/C inhibitors as promising therapeutic agents targeting cohesion-defective cancers.

SiO2 Nanoparticule-induced size-dependent genotoxicity – an in vitro study using sister chromatid exchange, micronucleus and comet assay

Abstract: Fine particles with a characteristic size smaller than 100 nm (ie nanoparticlesspread out in nowadays life Silicon or Si, is one of the most abundant chemical elements found on the Earth Its oxide forms, such as silicate (SiO4) and silicon dioxide, also known as silica (SiO2), are the main constituents of sand and quartz contributing to 90% of the Earth's crust In this work, three genotoxicity systems “sister chromatid exchange, cytokinesis block micronucleus test and single cell gel electrophoresis (comet) assay” were employed to provide further insight into the cytotoxic and mutagenic/genotoxic potential of SiO2 nanoparticules (particle size 6 nm, 20 nm, 50 nm) in cultured peripheral blood lymphocytes as in vitro It was observed that there is a significant decrease in Mitotic index (MI), Cytokinesis block proliferation index (CBPI), proliferation index (PRI) values expressed as Cell Kinetic parameters compared with negative control (p < 005) There is a statistically significant differenc

The effect of thiacloprid formulation on DNA/chromosome damage and changes in GST activity in bovine peripheral lymphocytes.

Abstract: The potential genotoxic effect of thiacloprid formulation on bovine peripheral lymphocytes was evaluated using the comet assay and the cytogenetic endpoints: chromosome aberrations (CAs), sister chromatid exchanges (SCEs) and micronuclei (MNi). Whole blood cultures were treated with the insecticide at concentrations of 30, 60, 120, 240 and 480 μg mL(-1) for 24, 48 h and/or 2 h of incubation. A statistically significant increase in the frequency of DNA damage, as well as in unstable chromosome aberrations (% breaks) were found after exposure to the insecticide at concentrations ranging from 120 to 480 μg mL(-1) (P < 0.05, P < 0.01, P < 0.001). For the detection of stable structural chromosome aberrations (e.g., translocations) and numerical aberrations by the FISH method, three whole chromosome painting probes for bovine chromosomes 1, 5 and 7 (BTA1, BTA5 and BTA7) were used in our experiments. We observed numerical aberrations, but without any statistical significance. Regarding the sister chromatid exchanges, no significant elevation in the SCE frequencies was found after 24-h exposure to the insecticide. A dose-related response in the SCE induction was obtained in bovine cultures after the prolonged time of exposure (48 h) to thiacloprid formulation at concentrations ranging from 120 to 480 μg mL(-1) in each donor (P < 0.05, P < 0.01), which was associated with a reduction of the PI (P < 0.05, P < 0.01). The insecticide failed to produce MNi; however, a significant reduction of CBPI was observed. Using real-time PCR, a decrease in the expression of bovine glutathione S-transferase M3 (GSTM3) was detected at the lowest dose. The higher concentrations of thiacloprid formulation caused an increase in the mRNA expression.

Defects in the Fanconi Anemia Pathway and Chromatid Cohesion in Head and Neck Cancer

Abstract: Failure to repair DNA damage or defective sister chromatid cohesion, a process essential for correct chromosome segregation, can be causative of chromosomal instability (CIN), which is a hallmark of many types of cancers. We investigated how frequent this occurs in head and neck squamous cell carcinoma (HNSCC) and whether specific mechanisms or genes could be linked to these phenotypes. The genomic instability syndrome Fanconi anemia is caused by mutations in any of at least 16 genes regulating DNA interstrand crosslink (ICL) repair. Since patients with Fanconi anemia have a high risk to develop HNSCC, we investigated whether and to which extent Fanconi anemia pathway inactivation underlies CIN in HNSCC of non-Fanconi anemia individuals. We observed ICL-induced chromosomal breakage in 9 of 17 (53%) HNSCC cell lines derived from patients without Fanconi anemia. In addition, defective sister chromatid cohesion was observed in five HNSCC cell lines. Inactivation of FANCM was responsible for chromosomal breakage in one cell line, whereas in two other cell lines, somatic mutations in PDS5A or STAG2 resulted in inadequate sister chromatid cohesion. In addition, FANCF methylation was found in one cell line by screening an additional panel of 39 HNSCC cell lines. Our data demonstrate that CIN in terms of ICL-induced chromosomal breakage and defective chromatid cohesion is frequently observed in HNSCC. Inactivation of known Fanconi anemia and chromatid cohesion genes does explain CIN in the minority of cases. These findings point to phenotypes that may be highly relevant in treatment response of HNSCC.

Detection of genotoxicity and mutagenicity of chlorthiophos using micronucleus, chromosome aberration, sister chromatid exchange, and Ames tests.

Abstract: Potential mutagenic and genotoxic effects of Chlorthiophos, an organophosphate pesticide, were evaluated using four standard assays. Five different concentrations of the pesticide were tested by an Ames test using Salmonella typhimurium strains TA97, TA98, TA100, and TA102, with and without S9 metabolic activation. No concentrations of Chlorthiophos showed mutagenic activity on the TA97, TA100, and TA102 strains, with and without S9 fraction, but were all mutagenic to the TA98 strain without S9. Sister chromatid exchange (SCE), chromosome aberration (CA), and micronucleus (MN) tests were used to investigate the genotoxic effects of Chlorthiophos in human peripheral lymphocytes treated with 25, 50, 100, and 200 µg/mL concentrations of Chlorthiophos for 24 and 48 h. The nuclear division index (NDI), replication index (RI), and mitotic index (MI) were also calculated to determine the cytotoxicity of Chlorthiophos. No increase in SCE frequency was seen for any treatment period or concentration, but Chlorthiophos at 200 µg/mL increased the frequency of CAs. Increases in MN formation were only observed at Chlorthiophos concentrations of 200 µg/mL following 24 and 48 h treatments. Chlorthiophos treatment reduced the MI and NDI significantly, but had no effect on the RI.

The in vitro genotoxic and cytotoxic effects of remeron on human peripheral blood lymphocytes.

Abstract: Remeron (Mirtazapine) is an antidepressant drug which exerts its action by blocking presynaptic α-2-adrenergic receptors and postsynaptic serotonin types 2 and 3 receptors. In this in vitro analysis, human peripheral blood lymphocytes was treated by remeron (10, 25, 40 and 55 μg/mL) for 24 hours and 48 hours periods, then it was attempted to study of genotoxic and cytotoxic effects of the substance on human peripheral blood lymphocytes by some tests such as sister chromatid exchange (SCE), chromosomal abnormalities (CA) and micronucleus (MN) tests. Also proliferating effect of the substance was investigated. Remeron didn’t significantly cause chromosomal abnormalities and sister chromatid exchange while caused micronucleus at 40 μg/mL concentration and 24 h periodic time and increased proliferation index of the both 24 and 48 hours treated cells was decreased in a concentration manner. Also, exposing to the remeron for 24 and 48 hours leaded to a decrease in mitotic index and nucleus division inde...

Assessment of genotoxicity associated with Behcet’s disease using sister-chromatid exchange assay: vitamin E versus mitomycin C

Abstract: Behcet’s disease (BD) is a multisystemic chronic inflammatory disorder that presents throughout the world with high frequency in Turkey and Middle East. BD has been shown to be associated with genotoxicity as patients with the disease have demonstrated high rates of sister chromatid exchange (SCE) and oxidative DNA damage. In this study, we examined the effect of vitamin E, which is known for its strong antioxidant activity, on the rate of SCE in cultured lymphocytes obtained from BD patients. In addition, the susceptibility of patient lymphocytes to the mutagenic agent mitomycin C (MMC) was also investigated. The results showed significant elevation in the rate of SCE in lymphocytes obtained from patients compared to those from healthy subjects (P < 0.01). Treatment with vitamin E normalized the elevated rate of SCE to a comparable level observed in the control group (P < 0.01). Finally, treatment of cultures with MMC significantly increased the rate of SCE in the lymphocytes of both patients and controls (P < 0.001). The magnitude of change in the rate of SCE induced by MMC was equivalent in both groups. This result suggests similar sensitivity of BD lymphocytes and control ones to MMC. In conclusion, genotoxicity associated with BD can be overcome by treatment with vitamin E. Lymphocytes of BD have normal sensitivity to the mutagenic agent MMC.

Investigation of cytotoxic and genotoxic effects of the antihistaminic drug, loratadine, on human lymphocytes

Abstract: Context: Loratadine (LOR) is a new generation antihistamine used in the treatment of allergic disorders. Objective: The aim of this study was to evaluate the cytogenotoxic effect of LOR on human peripheral blood lymphocytes. Materials and methods: We investigated the genotoxic effect of this drug in cultured human peripheral blood lymphocytes using sister chromatid exchange (SCE), chromosomal aberrations (CA) and micronucleus (MN) assay in culture conditions. Proliferation index (PI), mitotic index (MI) and nuclear division index (NDI) were also calculated to determine the cytotoxic/cytostatic effect. Cultures were treated with LOR at three concentrations (5, 15 and 25 µg/ml) for 48 h. Results: Although the MI significantly decreased at the higher concentrations (15 and 25 µg/ml) compared with negative (solvent) control, LOR indicated weaker cytotoxic potential in PI and NDI values at all the tested concentrations. LOR increased the frequencies of SCE, CA and MN in all lymphocyte cultures. However...

Can consumption of raw vegetables decrease the count of sister chromatid exchange? Results from a cross-sectional study in Krakow, Poland

Abstract: Background Sister chromatid exchange (SCE) is a widely used sensitive cytogenetic biomarker of exposure to genotoxic and cancerogenic agents Results of human monitoring studies and cytogenetic damage have revealed that biological effects of genotoxic exposures are influenced by confounding factors related to life-style Vegetable and fruit consumption may play a role, but available results are not consistent The purpose of the study was to investigate the effect of consumption of raw and cooked vegetables and fruits on SCE frequency

Association between sister chromatid exchange and double minute chromosomes in human tumor cells.

Abstract: Background Double minute chromosomes (DMs) are the cytogenetic hallmark of extra-chromosomal genomic amplification. They can well represent the advanced stage of malignancy. However, the mechanisms of DM generation are still not fully understood. Here, the sister chromatid exchange (SCE) was used to determine whether the occurrence of DMs was related to the high genomic instability in human carcinoma cells. We analyzed SCE frequencies in two groups of cell lines: the first group contained DM-positive cell lines such as UACC-1598, SK-PN-DW, and NCI-N87 carcinomas, while the second group comprised DM-negative cell lines including HO-8910, U251, and MGC-803.

Association of Polymorphisms of Phase I Metabolizing Genes with Sister Chromatid Exchanges in Occupational Workers Exposed to Toluene Used in Paint Thinners

Abstract: This study investigated genetic damage in paint workers mainly exposed to toluene as it is a major solvent used in paint thinners. Sister chromatid exchange (SCE) assay was used as biomarker of genotoxicity. Blood samples were collected from 30 paint workers and 30 control subjects matched with respect to age and other confounding factors except for exposure to toluene. SCE frequency was found to be significantly higher in paint workers (4.81 ± 0.92) as compared to control individuals (1.73 ± 0.54) (p < 0.05). We also investigated influence of polymorphisms of CYP2E1 and CYP1A1m2 genes on SCE frequency. Our results showed that there was significant increase in frequencies of SCE among the mutant genotypes of CYP2E1 and CYP1A1m2 as compared to wild genotypes. Our study indicated that long term exposure of toluene can increase genotoxic risk in paint workers.

Spontaneous sister chromatid exchange in mitotic chromosomes of the chinchilla (Chinchilla lanigera)

Abstract: Kuchta-Gladysz, M., Wojcik, E., Szeleszczuk, O., Niedbala, P. and Tyblewska, K. 2015. Spontaneous sister chromatid exchange in mitotic chromosomes of the chinchilla (Chinchilla lanigera). Can. J. Anim. Sci. 95: 543–550. The sister chromatid exchange (SCE) test is a cytogenetic tool with applications as a short-term screen. It is used to assess the influence of physical and chemical factors with potential mutagenic and genotoxic properties on the animal organism. The test results make it possible to eliminate mutagens, as well as helping to predict possible genetic consequences in animal cells and assess animal resistance. The mitotic chromosomes were obtained from an in vitro culture of peripheral blood lymphocytes with added bromodeoxyuridine (BrdU), at five different concentrations: 0.25, 0.5, 1.0, 2.5, and 5.0 µg mL−1. The chromosomes were stained according to the FPG method. Our analyses revealed the spontaneous SCE level in the chinchilla at the concentration of 0.5 µg mL−1. Higher concentrations of ...

In vitro genotoxicity and cytotoxicity of a particular combination of pemetrexed and cefixime in human peripheral blood lymphocytes

Abstract: This study aims to find the genotoxic and cytotoxic effects of a particular combination of pemetrexed (PMX) and cefixime (CFX) in human peripheral blood lymphocytes. Chromosome aberration (CA), sister chromatid exchange (SCE), and micronucleus (MN) tests were used to assess genotoxicity. Whereas, the cytotoxicity was evaluated by using mitotic index (MI), proliferation index (PI), and nuclear division index (NDI). Our tests were proceeded with concentrations of 12.5 + 450, 25 + 800, 37.5 + 1150, and 50 + 1500 μg/mL of a mixture of PMX and CFX separately for 24 hr and 48 hr. The combination of PMX + CFX did not induce the CA or SCE in human peripheral blood lymphocytes when compared with both the control and the solvent control. MN in human peripheral blood lymphocytes was not significantly increased after treatment with a particular combination of PMX + CFX. However, PMX + CFX significantly decreased the MI, PI and NDI at all concentrations for 24- and 48-hr treatment periods when compared with both controls. Generally, PMX + CFX inhibited cell proliferation more than positive control (MMC) and showed a higher cytotoxic effect than MMC at both treatment periods. These results were compared with individual effects of PMX and CFX. As a result, it was observed that a particular combination of PMX + CFX was not genotoxic. However, the combination synergistically increase cytotoxicity in human peripheral blood lymphocytes.

Genotoxic evaluation of terbinafine in human lymphocytes in vitro

Abstract: Terbinafine is an antimycotic drug usually used against several superficial fungal infections and with a potential application in the treatment of human cancers. Since to date there are few data on the genotoxic effects of terbinafine in mammalian cells, current study evaluated the potential genotoxic of such antifungal agent in cultured human peripheral blood lymphocytes. Terbinafine was used at the peak plasma concentration (1.0 μg/ml) and in four additional concentrations higher than the human plasmatic peak (5.0 μg/ml, 25.0 μg/ml, 50.0 μg/ml and 100.0 μg/ml). Chromosomal aberrations (CA), sister chromatid exchanges (SCE), micronuclei (MN), nucleoplasmic bridges (NP) and nuclear buds (NB) were scored as genetic endpoints. In all analysis no significant differences (α = 0.05, Kruskal-Wallis test) were observed. Complementary criterion adopted to obtain the final response in cytogenetic agreed with statistical results. Therefore, results of this study showed that terbinafine neither induced CA, SCE, MN, NP and NB nor affected significantly mitotic, replication and cytokinesis-block proliferation indices in any of the tested concentrations. It may be assumed that terbinafine was not genotoxic or cytotoxic to cultured human peripheral blood lymphocytes in our experimental conditions.

Genotoxicity of formaldehyde in peripheral blood lymphocytes among pathology laboratory workers

Abstract: Introduction: Formaldehyde (FA) is a chemical, traditionally used in pathology and anatomy laboratories as a tissue preservative. Many studies clearly indicated that FA can induce genotoxic effects in peripheral blood lymphocytes. Aim of work: to evaluate the genotoxic effects of formaldehyde among medical and paramedical personnel in The Histopathology Laboratory. Materials and Methods: The study was conducted in histopathology laboratory including 30 workers occupationally exposed to formaldehyde and 29 persons from other lab in kasr Al Aini hospital matching the exposed group for age, sex and socioeconomic status with no history of occupational exposure to formaldehyde. A questionnaire was done including inquiries about age, sex, occupational history and special habits. All participants were subjected to the following laboratory investigations: measuring DNA protein crosslink (DPC), sister chromatid exchange (SCE) and micronucleus (Mn) frequency in peripheral blood lymphocytes. Environmental monitoring of the work place was done to measure the level of FA at different place in the pathology lab. Results: This study showed statistically significant increase of DPC with significant increase in sister chromatid exchange (SCE) and micronucleus MN in peripheral blood lymphocytes. Conclusion: The studied group was exposed to formaldehyde level above the exposure limits recommended by NIOSH, ACGIH and OSHA with adverse health effects in the form of increase in DPC, SCE and Mn as genotoxicity biomarkers. These genotoxicity biomarkers are considered to play an important role in the carcinogenesis of FA.

Ispitivanje genetičke nestabilnosti primjenom in vitro analize izmjene sestrinskih kromatida

Abstract: Sister chromatid exchange from in vitro analysis (SCE) is a cytogenetic method used to control professionally exposed population by means of mutagenic or carcinogenic chemicals. It is the indirect indicator of the level of damage present in the DNA before duplication. The purpose was to determine if the exposure of xylene, methanol and acetic acid poses increased risk of DNA damage. The participants were people younger than 40 who do not work with chemicals or in a radiation zone. The analysis of SCE is based on the 72-hour cultivation of peripheral blood lymphocytes in the nutrient medium in the presence of 5-bromodeoxyuridine (BrdU) and phytohaemagglutinin (PHA). For each chemical, three different concentrations and control are tested. The results demonstrated that the chemicals cause an increase in the number of sister chromatid exchange in relation to the controls. There is a statistically significant reduction of mitotic index (P <0.05) at all concentrations of tested chemicals. This research demonstrates that all of tested chemicals cause an increased number of recombination repair in peripheral lymphocytes and have an antiproliferative effect associated with genotoxicity. Biomonitoring is an important part of health checkups of people occupationally exposed to health hazards. It is based on the measurement of biological indicators which indicate early biological effects that precede the onset of malignancies and other diseases. A routine analysis of SCE may indirectly show the quality of safety measures at a workplace.

Efectos genotóxicos del azinfos metílico en bacterias y cultivos de linfocitos humanos después de activación vegetal

Abstract: The evaluation of the potential risk of pesticides applied to crops consumed by humans in Mexico is appropriate and necessary because plant pro-mutagenic transformation in toxic metabolites and their subsequent incorporation involve a risk for health when such crops are ingested. Plant metabolism of agricultural insecticides produces compounds that could be introduced in the food chain, increasing the contamination and poisoning risk by agrochemical metabolism. In this study we evaluated the effect of the organophosphorus insecticide azinphos methyl transformed by S10 fraction of broad bean ( Vicia faba ), using as indicator of mutagenic damage the reverse mutation of Salmonella typhimurium strains TA98 and TA100 and the sister chromatid exchange (SCE) in human lymphocyte cultures. Results of mutagenicity showed that when Salmonella TA98 and TA100 strains were treated directly with azinphos methyl, negative response was obtained. The same occurred with human lymphocytes tested directly with this insecticide. When Vicia faba S10 enzymatic mix was added, there was a mutagenic response in both Salmonella strains. These results suggest that the mechanisms to induce mutations by azinphos methyl were frameshift mutation (TA98 strain) as well as pair bases substitution (TA100 strain). Likewise, SCE production was significant and dose-response relationship was observed in human lymphocyte cultures. The cell kinetics (M1, M2 and M3 cells), the replication index and the mitotic index are also analyzed. Only in the treatments with S10 fraction the effects were observed. At the highest concentration mitotic inhibition was produced.

The Association between Anti-Oxidant/Redox Status and Sister Chromatid Exchanges in Down Syndrome Individuals

Abstract: The main objective of this project is to study the possible association between anti-oxidant/redox status and DNA instability in Down syndrome. The activities of 5 antioxidant enzymes were studied in 19 Down syndrome (DS) cases and in age- and sex-matched normal controls. Sister Chromatid Exchanges (SCE) were measured in lymphocyte cultures derived from all DS and control subjects. All DS and control individuals had normal hematological parameters, but the proliferation and mitotic indices were significantly lower in the DS- than in the controls-derived lymphocyte cultures, while the average generation time was higher than that in the controls. The specific activity of superoxide dismutase in the DS individuals was 40% higher than that in the controls, while the specific activity of glutathione S-transferase in the DS group was significantly lower than that in the controls (P ≤ 0.05). Catalase and glutathione peroxides’ activities were not different between the two groups (P > 0.05). SCE rate in the DS derived cultures was significantly higher (P < 0.001) than that of the controls. DS individuals have a higher oxidative stress, higher superoxide dismutase activities and higher rates of SCE in their derived lymphocyte cultures compared to those of the controls. We claim that such differences may have resulted from the over expression of superoxide dismutase gene, leading to imbalanced cellular antioxidant mechanisms and, consequently, resulted in a high concentration of free radicals that destabilized the DNA as expressed by the high rate of SCE.

In vitro enhancement of doxorubicin genotoxic activities and interference with cell cycle delay by Plumbago indica root ethanolic extract in human lymphocytes.

Abstract: Background: Combinations of modern medicines with herbal medicines are being developed for more effectiveness. Data on the safety and drug-herb interactions are needed to be clarified. Ethanolic extract of Plumbago indica root (EEPIR) is medicinally used for cancer treatment in Asian traditional medicine. However, its mechanism of action is still inconclusive. Our previous study demonstrated that EEPIR was genotoxic and induced cell cycle delay in human lymphocytes in vitro. Objective: To investigate genotoxic potency and interference with cell cycle of EEPIR in combination with doxorubicin (DXR), a standard chemotherapeutic agent, in human lymphocytes by in vitro sister chromatid exchange (SCE) assay. Material and Method: Human lymphocytes were pretreated with EEPIR at 6.25-100 mcg/ml followed by DXR (0.1 mcg/ml). SCE levels and cell cycle kinetics were evaluated. Results: EEPIR pretreatments (6.5-50 mcg/ml) significantly enhanced genetic damage induced by DXR (p<0.05). Delaying of the cell cycle was detected and related to EEPIR concentration. EEPIR at 100 mcg/ml, on the contrary, did not enhance DXRinduced genotoxicity but tended to lower genotoxicity compared to DXR treatment alone. It significantly delayed cell cycle the most (p<0.05). Conclusion: EEPIR pretreatments at proper doses enhanced genotoxic damage induced by DXR in human lymphocytes. Delaying cell cycle by EEPIR could lower that potency. Usage of EEPIR, therefore, should be adjusted for safety. Combination of EEPIR with DXR might be useful for more efficient cancer treatment with less DXR toxicity. Further in vivo study is needed to support this in vitro evidence. Keywords: Plumbago indica root, Doxorubicin, Cell cycle, Sister chromatid exchange, Human lymphocytes, Genotoxicity