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https://www.readbyqxmd.com/read/28737676/high-dose-ascorbate-causes-both-genotoxic-and-metabolic-stress-in-glioma-cells
#1
Maria Leticia Castro, Georgia M Carson, Melanie J McConnell, Patries M Herst
We have previously shown that exposure to high dose ascorbate causes double stranded breaks (DSBs) and a build-up in S-phase in glioblastoma (GBM) cell lines. Here we investigated whether or not this was due to genotoxic stress as well as metabolic stress generated by exposure to high dose ascorbate, radiation, ascorbate plus radiation and H₂O₂ in established and primary GBM cell lines. Genotoxic stress was measured as phosphorylation of the variant histone protein, H2AX, 8-oxo-7,8-dihydroguanine (8OH-dG) positive cells and cells with comet tails...
July 22, 2017: Antioxidants (Basel, Switzerland)
https://www.readbyqxmd.com/read/28735740/dna-damage-and-the-bystander-response-in-tumor-and-normal-cells-exposed-to-x-rays
#2
M Subhashree, R Venkateswarlu, K Karthik, V Shangamithra, P Venkatachalam
Monolayer and suspension cultures of tumor (BMG-1, CCRF-CEM), normal (AG1522, HADF, lymphocytes) and ATM-mutant (GM4405) human cells were exposed to X-rays at doses used in radiotherapy (high dose and high dose-rate) or radiological imaging (low dose and low dose-rate). Radiation-induced DNA damage, its persistence, and possible bystander effects were evaluated, based on DNA damage markers (γ-H2AX, p53(ser15)) and cell-cycle-specific cyclins (cyclin B1 and cyclin D1). Dose-dependent DNA damage and a dose-independent bystander response were seen after exposure to high dose and high dose-rate radiation...
September 2017: Mutation Research
https://www.readbyqxmd.com/read/28732355/checkpoint-kinase-1-inhibition-sensitises-transformed-cells-to-dihydroorotate-dehydrogenase-inhibition
#3
Stéphanie Arnould, Geneviève Rodier, Gisèle Matar, Charles Vincent, Nelly Pirot, Yoann Delorme, Charlène Berthet, Yoan Buscail, Jean Yohan Noël, Simon Lachambre, Marta Jarlier, Florence Bernex, Hélène Delpech, Pierre Olivier Vidalain, Yves L Janin, Charles Theillet, Claude Sardet
Reduction in nucleotide pools through the inhibition of mitochondrial enzyme dihydroorotate dehydrogenase (DHODH) has been demonstrated to effectively reduce cancer cell proliferation and tumour growth. The current study sought to investigate whether this antiproliferative effect could be enhanced by combining Chk1 kinase inhibition. The pharmacological activity of DHODH inhibitor teriflunomide was more selective towards transformed mouse embryonic fibroblasts than their primary or immortalised counterparts, and this effect was amplified when cells were subsequently exposed to PF477736 Chk1 inhibitor...
July 12, 2017: Oncotarget
https://www.readbyqxmd.com/read/28724724/brct-domain-protein-brit1-influences-class-switch-recombination
#4
Wei-Feng Yen, Ashutosh Chaudhry, Bharat Vaidyanathan, William T Yewdell, Joseph N Pucella, Rahul Sharma, Yulong Liang, Kaiyi Li, Alexander Y Rudensky, Jayanta Chaudhuri
DNA double-strand breaks (DSBs) serve as obligatory intermediates for Ig heavy chain (Igh) class switch recombination (CSR). The mechanisms by which DSBs are resolved to promote long-range DNA end-joining while suppressing genomic instability inherently associated with DSBs are yet to be fully elucidated. Here, we use a targeted short-hairpin RNA screen in a B-cell lymphoma line to identify the BRCT-domain protein BRIT1 as an effector of CSR. We show that conditional genetic deletion of BRIT1 in mice leads to a marked increase in unrepaired Igh breaks and a significant reduction in CSR in ex vivo activated splenic B cells...
July 19, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28720507/formation-and-degradation-of-nitrogen-mustard-induced-mgmt-dna-crosslinking-in-16hbe-cells
#5
Jin Cheng, Feng Ye, Guorong Dan, Yuanpeng Zhao, Jiqing Zhao, Zhongmin Zou
N-methyl-2,2-di(chloroethyl)amine (HN2) is a kind of bifunctional alkyltating agent, which can react with nucleophilic groups in DNA and/or protein to form HN2-bridged crosslinking of target molecules, such as DNA-protein crosslinkings (DPC). O(6)-methylguanine-DNA methyltransferase (MGMT) is a DNA damage repair enzyme which solely repairs alkyl adduct on DNA directly. However, MGMT was detected to act as a protein cross-linked with DNA via alkylation in presence of HN2, and unexpectedly turned into a DNA damage enhancer in the form of MGMT-DNA cross-link (mDPC)...
July 15, 2017: Toxicology
https://www.readbyqxmd.com/read/28714949/aspartate-%C3%AE-hydroxylase-disrupts-mitochondrial-dna-stability-and-function-in-hepatocellular-carcinoma
#6
C Tang, Y Hou, H Wang, K Wang, H Xiang, X Wan, Y Xia, J Li, W Wei, S Xu, Z Lei, T M Pawlik, H Wang, M Wu, F Shen
The mechanism of aberrant mitochondrial genome and function in hepatocellular carcinoma (HCC) remains largely unknown. Our previous study demonstrated an increased expression of aspartate β-hydroxylase (ASPH) in HCC tissues, which was associated with tumor invasiveness and a worse prognosis. Currently, we unexpectedly observed the presence of ASPH in purified mitochondrial protein fraction. In addition, immunostaining of both exogenously and endogenously expressed ASPH showed a colocalization with mitochondrial biomarkers...
July 17, 2017: Oncogenesis
https://www.readbyqxmd.com/read/28714549/increased-level-of-dna-damage-in-some-organs-of-obese-zucker-rats-by-%C3%AE-h2ax-analysis
#7
Alessia Azzarà, Anna Chiaramonte, Erika Filomeni, Barbara Pinto, Stefano Mazzoni, Simona Piaggi, Maria Angela Guzzardi, Fabrizio Bruschi, Patricia Iozzo, Roberto Scarpato
In a recent study, we showed that lymphocytes of obese Italian children/adolescents displayed levels of double strand breaks (DSB), assayed as serine 139-phosphorylated histone H2AX (γ-H2AX), about eightfold higher than normal weight controls, and that 30% of this damage-generated micronuclei. These findings suggested that obese children could be at increased risk of obesity-mediated cancer later in life. We therefore aimed to assess the level of γ-H2AX in a genetic animal model of obesity (Zucker rat) to identify a genotoxic/carcinogenic risk in some organs...
July 17, 2017: Environmental and Molecular Mutagenesis
https://www.readbyqxmd.com/read/28714367/parp-1-serves-as-a-novel-molecular-marker-for-hepatocellular-carcinoma-in-a-southern-chinese-zhuang-population
#8
Jiatong Li, Dongwei Dou, Ping Li, Wenqi Luo, Wenxin Lv, Chengdong Zhang, Xiaowei Song, Yuan Yang, Yuening Zhang, Yanzhen Xu, Feifan Xiao, Yan Wei, Jian Qin, Hongtao Li, Xiaoli Yang
PARP-1 (poly(ADP-ribose) polymerase-1) plays an important role in tumorigenesis. Since its effects on different populations are varied, this study investigated the impact of PARP-1 on primary hepatocellular carcinoma in a Southern Chinese Zhuang population. We assessed the global PARP-1 messenger RNA expression in patients with hepatocellular carcinoma using The Cancer Genome Atlas dataset. Increased PARP-1 expression, related to alpha-fetoprotein level, was observed. The area under the receiver operating characteristic curve value was 0...
July 2017: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
https://www.readbyqxmd.com/read/28713773/the-cytolethal-distending-toxin-subunit-cdtb-of-helicobacter-hepaticus-promotes-senescence-and-endoreplication-in-xenograft-mouse-models-of-hepatic-and-intestinal-cell-lines
#9
Christelle Péré-Védrenne, Martina Prochazkova-Carlotti, Benoit Rousseau, Wencan He, Lucie Chambonnier, Elodie Sifré, Alice Buissonnière, Pierre Dubus, Francis Mégraud, Christine Varon, Armelle Ménard
Cytolethal distending toxins (CDTs) are common among pathogenic bacteria of the human and animal microbiota. CDTs exert cytopathic effets, via their active CdtB subunit. No clear description of those cytopathic effects has been reported at the cellular level in the target organs in vivo. In the present study, xenograft mouse models of colon and liver cell lines were set up to study the effects of the CdtB subunit of Helicobacter hepaticus. Conditional transgenic cell lines were established, validated in vitro and then engrafted into immunodeficient mice...
2017: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/28710765/immunofluorescence-analysis-of-%C3%AE-h2ax-foci-in-mammalian-fibroblasts-at-different-phases-of-the-cell-cycle
#10
Liudmila Solovjeva, Denis Firsanov, Nadezhda Pleskach, Maria Svetlova
H2AX phosphorylation at Ser139 (formation of γ-H2AX) is an indicator of double-strand breaks in DNA (DSBs) after the action of different genotoxic stresses, including ionizing radiation, environmental agents, and chemotherapy drugs. The sites of DSBs can be visualized as focal sites of γ-H2AX using antibodies and immunofluorescence microscopy. The microscopy technique is the most sensitive method of DSB detection in individual cells. It is useful for experimental research, radiation biodosimetry, and clinical practice...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28710759/rapid-detection-of-%C3%AE-h2ax-by-flow-cytometry-in-cultured-mammalian-cells
#11
Denis Firsanov, Liudmila Solovjeva, Olga Lublinskaya, Valeriy Zenin, Igor Kudryavtsev, Maria Serebryakova, Maria Svetlova
Methods commonly used for detection of DNA double-strand breaks (DSBs) and analysis of cell death are generally time-consuming, and, therefore, any improvements in these techniques are important for researchers and clinicians. At present, flow cytometry is the most rapid method for detection of DSBs and cell viability. In this chapter, we provide our experience and methodological modification of flow cytometry protocol for the detection of γ-H2AX, a well-known marker of DSBs, in fixed mammalian fibroblasts...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28710758/express-%C3%AE-h2ax-immunocytochemical-detection-of-dna-damage
#12
Nate Hopp, Jodi Hagen, Birte Aggeler, Alexander E Kalyuzhny
DNA can be damaged by many environmental factors including chemical agents and ionizing radiation which induce the formation of DNA double-stranded breaks (DSBs). If DSBs are not repaired in a timely fashion this may cause the disruption of genome integrity, which can result in cancer development. Typically, DSBs are followed by phosphorylation of histone protein H2AX, a member of the H2A family. Immunocytochemical detection of phosphorylated H2AX (e.g., γ-H2AX) appears to be a useful technique for assessing DNA damage...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28698766/apple-flavonoids-suppress-carcinogen-induced-dna-damage-in-normal-human-bronchial-epithelial-cells
#13
Vazhappilly Cijo George, H P Vasantha Rupasinghe
SCOPE: Human neoplastic transformation due to DNA damage poses an increasing global healthcare concern. Maintaining genomic integrity is crucial for avoiding tumor initiation and progression. The present study aimed to investigate the efficacy of an apple flavonoid fraction (AF4) against various carcinogen-induced toxicity in normal human bronchial epithelial cells and its mechanism of DNA damage response and repair processes. METHODS AND RESULTS: AF4-pretreated cells were exposed to nicotine-derived nitrosamine ketones (NNK), NNK acetate (NNK-Ae), methotrexate (MTX), and cisplatin to validate cytotoxicity, total reactive oxygen species, intracellular antioxidants, DNA fragmentation, and DNA tail damage...
2017: Oxidative Medicine and Cellular Longevity
https://www.readbyqxmd.com/read/28690315/targeting-nucleolin-for-better-survival-in-diffuse-large-b-cell-lymphoma
#14
N Jain, H Zhu, T Khashab, Q Ye, B George, R Mathur, R K Singh, Z Berkova, J F Wise, F K Braun, X Wang, K Patel, Z Y Xu-Monette, J Courty, K H Young, L Sehgal, F Samaniego
Anthracyclines have been a cornerstone in the cure of diffuse large B-cell lymphoma (DLBCL) and other hematological cancers. The ability of anthracyclines to eliminate DLBCL depends on the presence of topoisomerase-II-alpha (TopIIA), a DNA repair enzyme complex. We identified nucleolin as a novel binding partner of TopIIA. Abrogation of nucleolin sensitized DLBCL cells to TopIIA targeting agents (doxorubicin/etoposide). Silencing nucleolin and challenging DLBCL cells with doxorubicin enhanced the phosphorylation of H2AX (γH2AX-marker of DNA damage) and allowed DNA fragmentation...
July 10, 2017: Leukemia: Official Journal of the Leukemia Society of America, Leukemia Research Fund, U.K
https://www.readbyqxmd.com/read/28689138/enhanced-dna-double-strand-break-repair-of-microbeam-targeted-a549-lung-carcinoma-cells-by-adjacent-wi38-normal-lung-fibroblast-cells-via-bi-directional-signaling
#15
Alisa Kobayashi, Tengku Ahbrizal Farizal Tengku Ahmad, Narongchai Autsavapromporn, Masakazu Oikawa, Shino Homma-Takeda, Yoshiya Furusawa, Jun Wang, Teruaki Konishi
Understanding the mechanisms underlying the radiation-induced bystander effect (RIBE) and bi-directional signaling between irradiated carcinoma cells and their surrounding non-irradiated normal cells is relevant to cancer radiotherapy. The present study investigated propagation of RIBE signals between human lung carcinoma A549 cells and normal lung fibroblast WI38 cells in bystander cells, either directly or indirectly contacting irradiated A549 cells. We prepared A549-GFP/WI38 co-cultures and A549-GFP/A549 co-cultures, in which A549-GFP cells stably expressing H2BGFP were co-cultured with either A549 cells or WI38 cells, respectively...
June 23, 2017: Mutation Research
https://www.readbyqxmd.com/read/28688881/the-abdominal-skin-of-female-sprague-dawley-rats-is-more-sensitive-than-the-back-skin-to-drug-induced-phototoxicity
#16
Kazuhiro Kuga, Hironobu Yasuno, Yumi Sakai, Yumiko Harada, Fumi Shimizu, Yumiko Miyamoto, Yuki Takamatsu, Makoto Miyamoto, Keiichiro Sato
In vivo phototoxicity studies are important to predict drug-induced phototoxicity in humans; however, a standard methodology has not established. To determine differences in sensitivity to drug-induced phototoxicity among various skin sites, we evaluated phototoxic reactions in the back and abdominal skin of female Sprague-Dawley rats orally dosed with phototoxic drugs (pirfenidone, 8-methoxysoraren, doxycycline, and lomefloxacin) or a non-phototoxic drug (gatifloxacin) followed by solar-simulated light irradiation comprising 18J/cm(2) ultraviolet A...
July 5, 2017: Journal of Pharmacological and Toxicological Methods
https://www.readbyqxmd.com/read/28685056/pml-silencing-inhibits-cell-proliferation-and-induces-dna-damage-in-cultured-ovarian-cancer-cells
#17
Sheng-Bing Liu, Zhong-Fei Shen, Yan-Jun Guo, Li-Xian Cao, Ying Xu
The promyelocytic leukemia (PML) gene is a tumor suppressor gene. It was first identified in acute promyelocytic leukemia, in which it is fused to retinoic acid receptor α by the (15;17) chromosomal translocation. The function of the PML protein is frequently lost or aberrant in human solid tumors. In human ovarian carcinoma tissue, PML detected by immunohistochemistry was highly expressed. A PML-silencing vector, pSRG-shPml, was constructed and used to transfect human ovarian cancer cells. Cells were cultured and selected with puromycin for 10-15 days, and then the PML mRNA expression levels were detected by RT-qPCR and immunofluorescence...
July 2017: Biomedical Reports
https://www.readbyqxmd.com/read/28681635/estimation-of-low-dose-radiation-responsive-proteins-in-the-absence-of-genomic-instability-in-normal-human-fibroblast-cells
#18
Ji-Hye Yim, Jung Mi Yun, Ji Young Kim, Seon Young Nam, Cha Soon Kim
PURPOSE: Low-dose radiation has various biological effects such as adaptive responses, low-dose hypersensitivity, as well as beneficial effects. However, little is known about the particular proteins involved in these effects. Here, we sought to identify low-dose radiation-responsive phosphoproteins in normal fibroblast cells. MATERIALS AND METHODS: We assessed genomic instability and proliferation of fibroblast cells after γ-irradiation by γ-H2AX foci and micronucleus formation analyses and BrdU incorporation assay, respectively...
July 6, 2017: International Journal of Radiation Biology
https://www.readbyqxmd.com/read/28676400/inhibition-of-b7-h3-reverses-oxaliplatin-resistance-in-human-colorectal-cancer-cells
#19
Pengfei Zhang, Zhen Chen, Kuan Ning, Jian Jin, Xiaofeng Han
B7-H3, an immunoregulatory protein, has been found highly expressed in several cancer types, and involved in cancer cell migration and invasion. Here, we investigated the role of B7-H3 in oxaliplatin resistance in colorectal cancer (CRC) cells. Transient silencing of B7-H3 enhanced oxaliplatin sensitivity by increasing oxaliplatin-induced DNA damage. The overexpression of B7-H3 increased oxaliplatin resistance reducing the formation of phosphorylated histone H2AX (γH2AX) loci. The silencing of X-ray repair cross complementing group 1 (XRCC1), upregulated in B7-H3 overexpressing cells, induced an increase in cell death following oxaliplatin treatment...
July 1, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28676265/ultrafine-particles-ufps-from-domestic-wood-stoves-genotoxicity-in-human-lung-carcinoma-a549-cells
#20
Marabini Laura, Ozgen Senem, Turacchi Silvia, Aminti Stefania, Arnaboldi Francesca, Lonati Giovanni, Fermo Paola, Corbella Lorenza, Valli Gianluigi, Bernardoni Vera, Dell'Acqua Manuela, Vecchi Roberta, Becagli Silvia, Caruso Donatella, Corrado L Galli, Marinovich Marina
In this paper, results on the potential toxicity of ultrafine particles (UFPs d<100nm) emitted by the combustion of logwood and pellet (hardwood and softwood) are reported. The data were collected during the TOBICUP (TOxicity of BIomass COmbustion generated Ultrafine Particles) project, carried out by a team composed of interdisciplinary research groups. The genotoxic evaluation was performed on A549 cells (human lung carcinomacells) using UFPs whose chemical composition was assessed by a suite of analytical techniques...
August 2017: Mutation Research
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