RÉSUMÉ
African swine fever virus (ASFV), as a contagious viral pathogen, is responsible for the occurrence of African swine fever (ASF), a rapidly spreading and highly lethal disease. Since ASFV was introduced into China in 2018, it has been quickly spread to many provinces, which brought great challenges to the pig industry in China. Due to the limited knowledge about the pathogenesis of ASFV, neither vaccines nor antiviral drugs are available. We have found that ASFV infection can induce oxidative stress responses in cells, and DNA repair enzymes play a key role in this process. This study employed RNA interference, RT-qPCR, Western blotting, Hemadsorption (HAD), and flow cytometry to investigate the effects of the inhibitors of DNA repair enzymes OGG1 and MTH1 on ASFV replication and evaluated the anti-ASFV effects of the inhibitors. This study provides reference for the development of anti-viral drugs.
Sujet(s)
Virus de la peste porcine africaine , DNA Glycosylases , Phosphoric monoester hydrolases , Réplication virale , Virus de la peste porcine africaine/génétique , Virus de la peste porcine africaine/effets des médicaments et des substances chimiques , Animaux , Réplication virale/effets des médicaments et des substances chimiques , Suidae , DNA Glycosylases/métabolisme , DNA Glycosylases/génétique , Phosphoric monoester hydrolases/génétique , Phosphoric monoester hydrolases/antagonistes et inhibiteurs , Phosphoric monoester hydrolases/métabolisme , Enzymes de réparation de l'ADN/génétique , Enzymes de réparation de l'ADN/métabolisme , Peste porcine africaine/virologie , Antiviraux/pharmacologie , Interférence par ARN , Petit ARN interférent/génétique , Antienzymes/pharmacologie , Stress oxydatif/effets des médicaments et des substances chimiques , Cellules VeroRÉSUMÉ
Relatively low levels of antioxidant enzymes coupled with high oxygen metabolism result in the formation of numerous oxidative DNA damages in the tissues of the central nervous system. Recently, kynurenic acid (KYNA), knowns for its neuroprotective properties, has gained increasing attention in this context. Therefore, our hypothesis assumed that increased KYNA levels in the brain would positively influence mRNA expression of selected enzymes of the base excision repair pathway as well as enhance their efficiency in excising damaged nucleobases in specific areas of the sheep brain. The study was conducted on adult anestrous sheep (n = 18), in which two different doses of KYNA (20 and 100 µg/day) were infused into the third brain ventricle for three days. Molecular and biochemical analysis included the hypothalamus (preoptic and mediol-basal areas), hippocampus (CA3 field) and amygdala (central amygdaloid nucleus), dissected from the brain of sheep euthanized immediately after the last infusion. The results revealed a significant increase P < 0.001) in the relative mRNA abundance of N-methylpurine DNA glycosylase (MPG) following administration of both dose of KYNA across all examined tissues. The transcription of thymine-DNA glycosylase (TDG) increased significantly (P < 0.001) in all tissues in response to the lower KYNA dose compared to the control group. Moreover, 8-oxoguanine (8-oxoG) DNA glycosylase (OGG1) mRNA levels were also higher in both animal groups (P < 0.001). In addition, in the hypothalamus, hippocampus and amygdala, AP endonuclease 1 (APE1) mRNA expression increased under both doses of KYNA. Moreover, the both dose of KYNA significantly stimulated the efficiency of 8-oxoG excision in hypothalamus and amygdala (P < 0.05-0.001). The lower and higher doses of KYNA significantly influenced the effectiveness of εA and εC in all structures (P < 0.01-0.001). In conclusion, the favorable effect of KYNA in the brain may include the protection of genetic material in nerve and glial cells by stimulating the expression and efficiency of BER pathway enzymes.
Sujet(s)
Encéphale , DNA Glycosylases , Réparation de l'ADN , Acide kynurénique , Animaux , Réparation de l'ADN/effets des médicaments et des substances chimiques , Ovis , Acide kynurénique/métabolisme , DNA Glycosylases/métabolisme , DNA Glycosylases/génétique , Encéphale/métabolisme , Encéphale/effets des médicaments et des substances chimiques , Hypothalamus/métabolisme , Hypothalamus/effets des médicaments et des substances chimiques , ARN messager/métabolisme , ARN messager/génétique , Altération de l'ADN/effets des médicaments et des substances chimiques , DNA-(apurinic or apyrimidinic site) lyase/métabolisme , DNA-(apurinic or apyrimidinic site) lyase/génétique , Femelle , Hippocampe/métabolisme , Hippocampe/effets des médicaments et des substances chimiques , Réparation par excisionRÉSUMÉ
Maintaining the integrity of the genome is fundamental to living organisms. To this end, nature developed several mechanisms to find and promptly repair DNA lesions. Among them, base excision repair (BER) enzymes evolved to efficiently carry out this task. Notably, the mechanisms allowing these proteins to search for, detect, and fix DNA damage on a biologically relevant time scale still remain partially unclear. By taking MutY, a BER enzyme implied in the repair of the 8-oxoguanine-adenine mismatches, as a model system, we shed some light on the repair mechanism through a theoretical-computational approach. First, we estimated the effect of the oxidation state of the MutY iron-sulfur cluster on the protein-DNA binding. Then, the redox thermodynamics of both the protein cluster and DNA nucleobases are calculated. Finally, the charge migration kinetics along the double strand bound to the enzyme has been evaluated. The rationalization of our results indicates that the search for DNA lesions is essentially dictated by the redox chemistry of the species involved, i.e., the iron-sulfur redox cofactor and the DNA bound to the enzyme.
Sujet(s)
DNA Glycosylases , Réparation de l'ADN , Oxydoréduction , DNA Glycosylases/métabolisme , DNA Glycosylases/composition chimique , Réparation de l'ADN/physiologie , ADN/métabolisme , ADN/composition chimique , Cinétique , Altération de l'ADN , Thermodynamique , Ferrosulfoprotéines/métabolisme , Ferrosulfoprotéines/composition chimique , Ferrosulfoprotéines/génétiqueRÉSUMÉ
DNA glycosylases initiate the base excision repair (BER) pathway by catalyzing the removal of damaged or mismatched bases from DNA. The Arabidopsis DNA glycosylase methyl-CpG-binding domain protein 4 like (MBD4L) is a nuclear enzyme triggering BER in response to the genotoxic agents 5-fluorouracil and 5-bromouracil. To date, the involvement of MBD4L in plant physiological processes has not been analyzed. To address this, we studied the enzyme functions in seeds. We found that imbibition induced the MBD4L gene expression by generating two alternative transcripts, MBD4L.3 and MBD4L.4. Gene activation was stronger in aged than in non-aged seeds. Seeds from mbd4l-1 mutants displayed germination failures when maintained under control or ageing conditions, while 35S:MBD4L.3/mbd4l-1 and 35S:MBD4L.4/mbd4l-1 seeds reversed these phenotypes. Seed nuclear DNA repair, assessed by comet assays, was exacerbated in an MBD4L-dependent manner at 24 h post-imbibition. Under this condition, the BER genes ARP, APE1L, and LIG1 showed higher expression in 35S:MBD4L.3/mbd4l-1 and 35S:MBD4L.4/mbd4l-1 than in mbd4l-1 seeds, suggesting that these components could coordinate with MBD4L to repair damaged DNA bases in seeds. Interestingly, the ATM, ATR, BRCA1, RAD51, and WEE1 genes associated with the DNA damage response (DDR) pathway were activated in mbd4l-1, but not in 35S:MBD4L.3/mbd4l-1 or 35S:MBD4L.4/mbd4l-1 seeds. These results indicate that MBD4L is a key enzyme of a BER cascade that operates during seed imbibition, whose deficiency would cause genomic damage detected by DDR, generating a delay or reduction in germination.
Sujet(s)
Protéines d'Arabidopsis , Arabidopsis , DNA Glycosylases , Réparation de l'ADN , Germination , Graines , Graines/génétique , Graines/métabolisme , Arabidopsis/génétique , Arabidopsis/métabolisme , Protéines d'Arabidopsis/métabolisme , Protéines d'Arabidopsis/génétique , DNA Glycosylases/métabolisme , DNA Glycosylases/génétique , Régulation de l'expression des gènes végétaux , Altération de l'ADNRÉSUMÉ
Objectives: Exploring the relationship between the hOGG1 rs1052133 polymorphism and the occurrence of nasopharyngeal carcinoma (NPC). Methods: PubMed, Web of Science, Scopus, CNKI, Wanfangdata, and VIP were used to search for studies and the NOS evaluation scale was used to evaluate the quality. All studies were grouped according to different genotypes. The Cochrane's Q test and I2 test were used for heterogeneity evaluations. If heterogeneity was small, the fixed effects model was used, and conversely, the random effects model was used. Publication bias was also detected. P < .05 in all results indicated statistically significant. Results: We ultimately included 6 studies with 2021 NPC patients in the study group and 2375 healthy populations in the control group. After meta-analysis, it was found that the total OR value of the "Ser/Cys (CG) vs Ser/Ser (CC)" group was 1.00 (95% CI: 0.85-1.18) and the "Cys/Cys (GG) vs Ser/Ser (CC)" group was 1.06 (95% CI: 0.87-1.28). These results were not statistically significant (P > .05). Furthermore, the integrated total OR values of each group were not statistically significant with or without the smoking history, even in other genotype models (Allele, Dominant, Recessive, and Additive) (P > .05). Conclusion: There is no clear correlation between the hOGG1 rs1052133 polymorphism and the occurrence of NPC, even with or without the smoking history.
Sujet(s)
Allèles , DNA Glycosylases , Prédisposition génétique à une maladie , Génotype , Cancer du nasopharynx , Polymorphisme de nucléotide simple , Humains , Cancer du nasopharynx/génétique , DNA Glycosylases/génétique , Tumeurs du rhinopharynx/génétique , Odds ratio , Études d'associations génétiques , Biais de publication , Études cas-témoinsRÉSUMÉ
Maintenance of DNA integrity is essential to all forms of life. DNA damage generated by reaction with genotoxic chemicals results in deleterious mutations, genome instability, and cell death. Pathogenic bacteria encounter several genotoxic agents during infection. In keeping with this, the loss of DNA repair networks results in virulence attenuation in several bacterial species. Interstrand DNA crosslinks (ICLs) are a type of DNA lesion formed by covalent linkage of opposing DNA strands and are particularly toxic as they interfere with replication and transcription. Bacteria have evolved specialized DNA glycosylases that unhook ICLs, thereby initiating their repair. In this study, we describe AlkX, a DNA glycosylase encoded by the multidrug resistant pathogen Acinetobacter baumannii. AlkX exhibits ICL unhooking activity similar to that of its Escherichia coli homolog YcaQ. Interrogation of the in vivo role of AlkX revealed that its loss sensitizes cells to DNA crosslinking and impairs A. baumannii colonization of the lungs and dissemination to distal tissues during pneumonia. These results suggest that AlkX participates in A. baumannii pathogenesis and protects the bacterium from stress conditions encountered in vivo. Consistent with this, we found that acidic pH, an environment encountered during host colonization, results in A. baumannii DNA damage and that alkX is induced by, and contributes to, defense against acidic conditions. Collectively, these studies reveal functions for a recently described class of proteins encoded in a broad range of pathogenic bacterial species.
Sujet(s)
Acinetobacter baumannii , Altération de l'ADN , DNA Glycosylases , Acinetobacter baumannii/pathogénicité , Acinetobacter baumannii/génétique , Acinetobacter baumannii/enzymologie , Acinetobacter baumannii/métabolisme , DNA Glycosylases/métabolisme , DNA Glycosylases/génétique , Réparation de l'ADN , Infections à Acinetobacter/microbiologie , Infections à Acinetobacter/anatomopathologie , Protéines bactériennes/métabolisme , Protéines bactériennes/génétique , Animaux , Souris , ADN bactérien/génétique , ADN bactérien/métabolisme , Virulence , Escherichia coli/génétique , Escherichia coli/métabolismeRÉSUMÉ
CTC1-STN1-TEN1 (CST) is a single-stranded DNA binding protein vital for telomere length maintenance with additional genome-wide roles in DNA replication and repair. While CST was previously shown to function in double-strand break repair and promote replication restart, it is currently unclear whether it has specialized roles in other DNA repair pathways. Proper and efficient repair of DNA is critical to protecting genome integrity. Telomeres and other G-rich regions are strongly predisposed to oxidative DNA damage in the form of 8-oxoguanines, which are typically repaired by the base-excision repair (BER) pathway. Moreover, recent studies suggest that CST functions in the repair of oxidative DNA lesions. Therefore, we tested whether CST interacts with and regulates BER protein activity. Here, we show that CST robustly stimulates proteins involved in BER, including OGG1, Pol ß, APE1, and LIGI, on both telomeric and non-telomeric DNA substrates. Biochemical reconstitution of the pathway indicates that CST stimulates BER. Finally, knockout of STN1 or CTC1 leads to increased levels of 8-oxoguanine, suggesting defective BER in the absence of CST. Combined, our results define an undiscovered function of CST in BER, where it acts as a stimulatory factor to promote efficient genome-wide oxidative repair.
Sujet(s)
Altération de l'ADN , Réparation de l'ADN , Protéines télomériques , Humains , Protéines télomériques/métabolisme , Protéines télomériques/génétique , Télomère/métabolisme , Télomère/génétique , DNA Glycosylases/métabolisme , DNA Glycosylases/génétique , Stress oxydatif , DNA-(apurinic or apyrimidinic site) lyase/métabolisme , DNA-(apurinic or apyrimidinic site) lyase/génétique , Guanine/analogues et dérivés , Guanine/métabolisme , DNA polymerase beta/métabolisme , DNA polymerase beta/génétique , Réparation par excisionRÉSUMÉ
AIM: To identify mtDNA and OGG1 as potential biomarker candidates for mechanical asphyxia. METHOD: The human tissues are divided into experimental group (hanging and strangulation) and control groups (hemorrhagic shock, brain injury group, and poisoning group). Detected the expression of OGG1 and integrity of mtDNA in cardiac tissue of each group. We used over-OGG1 vector and siRNA-OGG1 transfecting H9C2 cell line to observe the function of OGG1 in hypoxic cells. RESULTS: 1. mtDNA integrity decreased in the mechanical asphyxia group, OGG1 expression increased in mechanical asphyxia groups. They can be biomarkers for mechanical asphyxia. 2. OGG1 increased first and decreased in hypoxia-induced H9C2 cells. OGG1 upregulated the TFAM, NRF1, and Bcl2 in hypoxia-induced H9C2. OGG1 downregulated cleaved-Caspase3 in hypoxia-induced H9C2 cells. 3. In the normoxia condition, NAC maintained mtDNA integrity and decreased the mitochondrial membrane potential and amount of ATP. CONCLUSION: mtDNA integrity and OGG1 expression can be biomarkers for mechanical asphyxia. OGG1 can maintain mtDNA integrity and maintain the stability of the mitochondrial membrane.
Sujet(s)
Asphyxie , Marqueurs biologiques , DNA Glycosylases , ADN mitochondrial , DNA Glycosylases/métabolisme , Humains , ADN mitochondrial/métabolisme , Marqueurs biologiques/métabolisme , Asphyxie/métabolisme , Lignée cellulaire , Potentiel de membrane mitochondriale , Animaux , Myocarde/métabolisme , MâleRÉSUMÉ
Two guanine base editors created using an engineered N-methylpurine DNA glycosylase with CRISPR systems achieved targeted G-to-T editing with 4.94-12.50% efficiency in rice (Oryza sativa). The combined use of the DNA glycosylase and deaminases enabled co-editing of target guanines with adenines or cytosines.
Sujet(s)
Édition de gène , Guanine , Oryza , Oryza/génétique , Édition de gène/méthodes , Guanine/métabolisme , Systèmes CRISPR-Cas/génétique , DNA Glycosylases/métabolisme , DNA Glycosylases/génétique , Thymine/métabolismeRÉSUMÉ
BACKGROUND: MUTYH germline monoallelic variants have been detected in a number of patients affected by breast/ovarian cancer or endometrial cancer, suggesting a potential susceptibility role, though their significance remains elusive since the disease mechanism is normally recessive. Hence, the aim of this research was to explore the hypothesis that a second hit could have arisen in the other allele in the tumor tissue. METHODS: we used Sanger sequencing and immunohistochemistry to search for a second MUTYH variant in the tumoral DNA and to assess protein expression, respectively. RESULTS: we detected one variant of unknown significance, one variant with conflicting interpretation of pathogenicity and three benign/likely benign variants; the MUTYH protein was not detected in the tumor tissue of half of the patients, and in others, its expression was reduced. CONCLUSIONS: our results fail to demonstrate that germinal monoallelic MUTYH variants increase cancer risk through a LOH (loss of heterozygosity) mechanism in the somatic tissue; however, the absence or partial loss of the MUTYH protein in many tumors suggests its dysregulation regardless of MUTYH genetic status.
Sujet(s)
Tumeurs du sein , DNA Glycosylases , Tumeurs de l'endomètre , Tumeurs de l'ovaire , Humains , DNA Glycosylases/génétique , Femelle , Tumeurs de l'endomètre/génétique , Tumeurs de l'endomètre/anatomopathologie , Tumeurs de l'ovaire/génétique , Tumeurs de l'ovaire/anatomopathologie , Tumeurs du sein/génétique , Tumeurs du sein/anatomopathologie , Adulte d'âge moyen , Perte d'hétérozygotie , Prédisposition génétique à une maladie , Sujet âgé , AdulteRÉSUMÉ
BACKGROUND: 8-Oxoguanine DNA glycosylase (OGG1), a well-known DNA repair enzyme, has been demonstrated to promote lung fibrosis, while the specific regulatory mechanism of OGG1 during pulmonary fibrosis remains unclarified. METHODS: A bleomycin (BLM)-induced mouse pulmonary fibrosis model was established, and TH5487 (the small molecule OGG1 inhibitor) and Mitochondrial division inhibitor 1 (Mdivi-1) were used for administration. Histopathological injury of the lung tissues was assessed. The profibrotic factors and oxidative stress-related factors were examined using the commercial kits. Western blot was used to examine protein expression and immunofluorescence analysis was conducted to assess macrophages polarization and autophagy. The conditional medium from M2 macrophages was harvested and added to HFL-1 cells for culture to simulate the immune microenvironment around fibroblasts during pulmonary fibrosis. Subsequently, the loss- and gain-of function experiments were conducted to further confirm the molecular mechanism of OGG1/PINK1. RESULTS: In BLM-induced pulmonary fibrosis, OGG1 was upregulated while PINK1/Parkin was downregulated. Macrophages were activated and polarized to M2 phenotype. TH5487 administration effectively mitigated pulmonary fibrosis, M2 macrophage polarization, oxidative stress and mitochondrial dysfunction while promoted PINK1/Parkin-mediated mitophagy in lung tissues of BLM-induced mice, which was partly hindered by Mdivi-1. PINK1 overexpression restricted M2 macrophages-induced oxidative stress, mitochondrial dysfunction and mitophagy inactivation in lung fibroblast cells, and OGG1 knockdown could promote PINK1/Parkin expression and alleviate M2 macrophages-induced mitochondrial dysfunction in HFL-1 cells. CONCLUSION: OGG1 inhibition protects against pulmonary fibrosis, which is partly via activating PINK1/Parkin-mediated mitophagy and retarding M2 macrophage polarization, providing a therapeutic target for pulmonary fibrosis.
Sujet(s)
Bléomycine , DNA Glycosylases , Modèles animaux de maladie humaine , Macrophages , Mitophagie , Protein kinases , Fibrose pulmonaire , Animaux , Mitophagie/effets des médicaments et des substances chimiques , Fibrose pulmonaire/métabolisme , Fibrose pulmonaire/induit chimiquement , Fibrose pulmonaire/étiologie , Fibrose pulmonaire/anatomopathologie , DNA Glycosylases/métabolisme , DNA Glycosylases/génétique , Souris , Macrophages/métabolisme , Protein kinases/métabolisme , Bléomycine/effets indésirables , Mâle , Ubiquitin-protein ligases/métabolisme , Ubiquitin-protein ligases/génétique , Stress oxydatif/effets des médicaments et des substances chimiques , Souris de lignée C57BL , Activation des macrophages , Humains , QuinazolinonesRÉSUMÉ
OBJECTIVE: DNA repair genes and their variants have been found to alter the risk of oral cancer. METHOD: The level of expression of XRCC3, NBS1, and OGG1 genes among 20 cases of oral cancer, 6 pre-oral cancer, and 50 healthy control subjects was measured with RT-PCR. All the subjects were also genotyped for XRCC3 rs861539 C>T, NBS1 rs1805794 C>G, and OGG1 rs1052133 C>G polymorphisms by the PCR-RFLP method; their genotypes were correlated with their level of expression. Further, a localized fold structure analysis of the mRNA sequence surrounding the studied SNPs was performed with RNAfold. RESULTS: Results showed increased expression of XRCC3, NBS1, and OGG1 transcripts among oral cancer (4.49 fold, 3.45 fold, and 3.27 fold) as well as pre-oral cancer (3.04 fold, 5.32 fold, and 1.74 fold) as compared to control subjects. The transcript level of OGG1 was found to be significantly increased (6.68 fold, p-value 0.009) with the GG genotype compared to the CC genotype. The C>T polymorphism of XRCC3 and the C>G polymorphism of OGG1 result in an apparent change in its mRNA secondary structure. Folding energy with the C allele for XRCC3 C>T polymorphism was lower than that of the T allele (MFE C vs T: -50.20 kcal/mol vs -48.70 kcal/mol). In the case of OGG1 C>G polymorphism MFE for the C allele was higher (-23.30 kcal/mole) than with the G allele (-24.80 kcal/mol). CONCLUSION: Our results showed elevated levels of XRCC3, NBS1, and OGG1 both in oral cancer and pre-oral cancer conditions, which indicates their role as prospective biomarkers of oral cancer and pre-cancerous lesions. SNPs in these genes alter their level of expression, possibly by altering the secondary structure of their transcript. However, due to the small sample size our study can only provide a suggestive conclusion and warned future study with large sample size to verify our findings.
Sujet(s)
Marqueurs biologiques tumoraux , Protéines du cycle cellulaire , DNA Glycosylases , Réparation de l'ADN , Protéines de liaison à l'ADN , Tumeurs de la bouche , Protéines nucléaires , Polymorphisme de nucléotide simple , Humains , Tumeurs de la bouche/génétique , Tumeurs de la bouche/anatomopathologie , DNA Glycosylases/génétique , Marqueurs biologiques tumoraux/génétique , Mâle , Réparation de l'ADN/génétique , Études cas-témoins , Adulte d'âge moyen , Protéines de liaison à l'ADN/génétique , Femelle , Protéines nucléaires/génétique , Protéines du cycle cellulaire/génétique , Génotype , Carcinome épidermoïde/génétique , Carcinome épidermoïde/anatomopathologie , Adulte , ARN messager/génétique , Prédisposition génétique à une maladieRÉSUMÉ
BACKGROUND: Hereditary adenomatous polyposis syndromes, including familial adenomatous polyposis and other rare adenomatous polyposis syndromes, increase the lifetime risk of colorectal and other cancers. METHODS: A team of 38 experts convened to update the 2008 European recommendations for the clinical management of patients with adenomatous polyposis syndromes. Additionally, other rare monogenic adenomatous polyposis syndromes were reviewed and added. Eighty-nine clinically relevant questions were answered after a systematic review of the existing literature with grading of the evidence according to Grading of Recommendations, Assessment, Development, and Evaluation methodology. Two levels of consensus were identified: consensus threshold (≥67% of voting guideline committee members voting either 'Strongly agree' or 'Agree' during the Delphi rounds) and high threshold (consensus ≥ 80%). RESULTS: One hundred and forty statements reached a high level of consensus concerning the management of hereditary adenomatous polyposis syndromes. CONCLUSION: These updated guidelines provide current, comprehensive, and evidence-based practical recommendations for the management of surveillance and treatment of familial adenomatous polyposis patients, encompassing additionally MUTYH-associated polyposis, gastric adenocarcinoma and proximal polyposis of the stomach and other recently identified polyposis syndromes based on pathogenic variants in other genes than APC or MUTYH. Due to the rarity of these diseases, patients should be managed at specialized centres.
Sujet(s)
Adénocarcinome , Polypose adénomateuse colique , DNA Glycosylases , Tumeurs de l'estomac , Humains , Polypose adénomateuse colique/génétique , Polypose adénomateuse colique/thérapie , Polypose adénomateuse colique/diagnostic , Tumeurs de l'estomac/génétique , Tumeurs de l'estomac/thérapie , Tumeurs de l'estomac/diagnostic , Adénocarcinome/génétique , Adénocarcinome/thérapie , Adénocarcinome/diagnostic , DNA Glycosylases/génétique , Syndromes néoplasiques héréditaires/génétique , Syndromes néoplasiques héréditaires/thérapie , Syndromes néoplasiques héréditaires/diagnostic , Europe , Polypes adénomateux/génétique , Polypes adénomateux/thérapie , PolypesRÉSUMÉ
Ivermectin (IVM) is an anti-parasitic drug which is used for treating parasitic infestations. It has been used in humans for treating intestinal strongyloidiasis and onchocerciasis however, currently researchers are investigating its potential for treating coronavirus SARS-CoV-2. Due to its broad-spectrum activities, IVM is being used excessively in animals which has generated an interest for researchers to investigate its toxic effects. Cytotoxic and genotoxic effects have been reported in animals due to excessive usage of IVM. Therefore, this study aims to evaluate the cytotoxic and genotoxic effects of IVM on the Madin-Darby-Bovine-Kidney (MDBK) cell line by examining the expression of a DNA damage-responsive gene (OGG1). Cytotoxicity of IVM was tested using an assay (MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), whereas the genotoxicity was evaluated using comet assay along with micronucleus assay. Moreover, the gene expression of DNA damage response gene (OGG1) was measured by qRT-PCR, after extraction of RNA from the MDBK cell line using the TRIzol method and its conversion to cDNA by reverse-transcriptase PCR. During the experiment, cell viability percentage was measured at different doses of IVM i.e., 25%, 50%, 75%, along with LC50/2, LC50 and LC50*2. It was observed that the gene expression of OGG1 increased as the concentration of IVM increased. It was concluded that IVM has both cytotoxic and genotoxic effects on the MDBK cell line. Furthermore, it is recommended that studies related to the toxic effects of IVM at molecular level and on other model organisms should be conducted to combat its hazardous effects.
Sujet(s)
Altération de l'ADN , Ivermectine , Ivermectine/toxicité , Ivermectine/pharmacologie , Animaux , Altération de l'ADN/effets des médicaments et des substances chimiques , Lignée cellulaire , Bovins , Survie cellulaire/effets des médicaments et des substances chimiques , Tests de micronucleus , DNA Glycosylases/génétique , DNA Glycosylases/métabolisme , Test des comètes , Mutagènes/toxicité , Antiparasitaires/pharmacologie , Antiparasitaires/toxicité , Rein/effets des médicaments et des substances chimiques , Rein/cytologieRÉSUMÉ
The base excision repair (BER) pathway is a precise and versatile mechanism of DNA repair that is initiated by DNA glycosylases. Endonuclease VIII-like 1 (NEIL1) is a bifunctional glycosylase/abasic site (AP) lyase that excises a damaged base and subsequently cleaves the phosphodiester backbone. NEIL1 is able to recognize and hydrolyze a broad range of oxidatively-induced base lesions and substituted ring-fragmented guanines, including aflatoxin-induced 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl-formamido)-9-hydroxyaflatoxin B1 (AFB1-FapyGua). Due to NEIL1's protective role against these and other pro-mutagenic lesions, it was hypothesized that naturally occurring single nucleotide polymorphic (SNP) variants of NEIL1 could increase human risk for aflatoxin-induced hepatocellular carcinoma (HCC). Given that populations in South Asia experience high levels of dietary aflatoxin exposures and hepatitis B viral infections that induce oxidative stress, investigations on SNP variants of NEIL1 that occur in this region may have clinical implications. In this study, the most common South Asian variants of NEIL1 were expressed, purified, and functionally characterized. All tested variants exhibited activities and substrate specificities similar to wild type (wt)-NEIL1 on high-molecular weight DNA containing an array of oxidatively-induced base lesions. On short oligodeoxynucleotides (17-mers) containing either a site-specific apurinic/apyrimidinic (AP) site, thymine glycol (ThyGly), or AFB1-FapyGua, P206L-NEIL1 was catalytically comparable to wt-NEIL1, while the activities of NEIL1 variants Q67K and T278I on these substrates were ≈2-fold reduced. Variant T103A had a greatly diminished ability to bind to 17-mer DNAs, limiting the subsequent glycosylase and lyase reactions. Consistent with this observation, the rate of excision by T103A on 17-mer oligodeoxynucleotides containing ThyGly or AFB1-FapyGua could not be measured. However, the ability of T103A to excise ThyGly was improved on longer oligodeoxynucleotides (51-mers), with ≈7-fold reduced activity compared to wt-NEIL1. Our studies suggest that NEIL1 variant T103A may present a pathogenic phenotype that is limited in damage recognition, potentially increasing human risk for HCC.
Sujet(s)
DNA Glycosylases , Réparation de l'ADN , Polymorphisme de nucléotide simple , DNA Glycosylases/métabolisme , DNA Glycosylases/génétique , DNA Glycosylases/composition chimique , Humains , Aflatoxine B1/métabolisme , Altération de l'ADN , Carcinome hépatocellulaire/génétique , Carcinome hépatocellulaire/enzymologie , Spécificité du substrat , Tumeurs du foie/génétique , Tumeurs du foie/enzymologieRÉSUMÉ
Previous evidence suggests elevated levels of oxidatively-induced DNA damage, particularly 8-hydroxy-2'-deoxyguanosine (8-OH-dG), and abnormalities in the repair of 8-OH-dG by the base excision repair (BER) in bipolar disorder (BD). However, the genetic disposition of these abnormalities remains unknown. In this study, we aimed to investigate the levels of oxidatively-induced DNA damage and BER mechanisms in individuals with BD and their siblings, as compared to healthy controls (HCs). 46 individuals with BD, 41 siblings of individuals with BD, and 51 HCs were included in the study. Liquid chromatography-tandem mass spectrometry was employed to evaluate the levels of 8-OH-dG in urine, which were then normalized based on urine creatinine levels. The real-time-polymerase chain reaction was used to measure the expression levels of 8-oxoguanine DNA glycosylase 1 (OGG1), apurinic/apyrimidinic endonuclease 1 (APE1), poly ADP-ribose polymerase 1 (PARP1), and DNA polymerase beta (POLß). The levels of 8-OH-dG were found to be elevated in both individuals with BD and their siblings when compared to the HCs. The OGG1 and APE1 expressions were downregulated, while POLß expressions were upregulated in both the patient and sibling groups compared to the HCs. Age, smoking status, and the number of depressive episodes had an impact on APE1 expression levels in the patient group while body mass index, smoking status, and past psychiatric history had an impact on 8-OH-dG levels in siblings. Both individuals with BD and unaffected siblings presented similar abnormalities regarding oxidatively-induced DNA damage and BER, suggesting a link between abnormalities in DNA damage/BER mechanisms and familial susceptibility to BD. Our findings suggest that targeting the oxidatively-induced DNA damage and BER pathway could offer promising therapeutic strategies for reducing the risk of age-related diseases and comorbidities in individuals with a genetic predisposition to BD.
Sujet(s)
8-Hydroxy-2'-désoxyguanosine , Trouble bipolaire , Altération de l'ADN , DNA Glycosylases , Réparation de l'ADN , Stress oxydatif , Fratrie , Humains , Trouble bipolaire/génétique , Trouble bipolaire/métabolisme , Femelle , Mâle , Adulte , DNA Glycosylases/génétique , Stress oxydatif/génétique , Adulte d'âge moyen , DNA polymerase beta/génétique , DNA polymerase beta/métabolisme , DNA-(apurinic or apyrimidinic site) lyase/génétique , Études cas-témoins , Jeune adulte , Désoxyguanosine/analogues et dérivés , Désoxyguanosine/urine , Réparation par excisionRÉSUMÉ
Over the past decades, cancer stem cells (CSCs) have emerged as a critical subset of tumor cells associated with tumor recurrence and resistance to chemotherapy. Understanding the mechanisms underlying CSC-mediated chemoresistance is imperative for improving cancer therapy outcomes. This study delves into the regulatory role of NEIL1, a DNA glycosylase, in chemoresistance in ovarian CSCs. We first observed a decreased expression of NEIL1 in ovarian CSCs, suggesting its potential involvement in CSC regulation. Using pan-cancer analysis, we confirmed the diminished NEIL1 expression in ovarian tumors compared to normal tissues. Furthermore, NEIL1 downregulation correlated with an increase in stemness markers and enrichment of CSCs, highlighting its role in modulating CSC phenotype. Further mechanistic investigation revealed an inverse correlation between NEIL1 and RAD18 expression in ovarian CSCs. NEIL1 depletion led to heightened RAD18 expression, promoting chemoresistance possibly via enhancing Translesion DNA Synthesis (TLS)-mediated DNA lesion bypass. Moreover, dowregulation of NEIL1 results in reduced DNA damage accumulation and suppressed apoptosis in ovarian cancer. Overall, our findings unveil a novel mechanism involving NEIL1 and RAD18 in regulating chemoresistance in ovarian CSCs. Targeting this NEIL1-RAD18 axis may offer promising therapeutic strategies for combating chemoresistance and improving ovarian cancer treatment outcomes.
Sujet(s)
DNA Glycosylases , Protéines de liaison à l'ADN , Résistance aux médicaments antinéoplasiques , Cellules souches tumorales , Tumeurs de l'ovaire , Régulation positive , Humains , Femelle , Tumeurs de l'ovaire/métabolisme , Tumeurs de l'ovaire/anatomopathologie , Tumeurs de l'ovaire/génétique , Tumeurs de l'ovaire/traitement médicamenteux , Résistance aux médicaments antinéoplasiques/génétique , Cellules souches tumorales/métabolisme , Cellules souches tumorales/anatomopathologie , DNA Glycosylases/métabolisme , DNA Glycosylases/génétique , Lignée cellulaire tumorale , Protéines de liaison à l'ADN/métabolisme , Protéines de liaison à l'ADN/génétique , Régulation de l'expression des gènes tumoraux , Ubiquitin-protein ligases/génétique , Ubiquitin-protein ligases/métabolisme , Altération de l'ADN , Apoptose/effets des médicaments et des substances chimiques , Apoptose/génétiqueRÉSUMÉ
Cervical cancer cells possess high levels of reactive oxygen species (ROS); thus, increasing oxidative stress above the toxicity threshold to induce cell death is a promising chemotherapeutic strategy. However, the underlying mechanisms of cell death are elusive, and efficacy and toxicity issues remain. Within DNA, 8-oxo-7,8-dihydroguanine (8-oxoG) is the most frequent base lesion repaired by 8-oxoguanine glycosylase 1 (OGG1)-initiated base excision repair. Cancer cells also express high levels of MutT homolog 1 (MTH1), which prevents DNA replication-induced incorporation of 8-oxoG into the genome by hydrolyzing 8-oxo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP). Here, we revealed that ROS-inducing agents triggered cervical cancer to undergo parthanatos, which was mainly induced by massive DNA strand breaks resulting from overwhelming 8-oxoG excision by OGG1. Furthermore, the MTH1 inhibitor synergized with a relatively low dose of ROS-inducing agents by enhancing 8-oxoG loading in the DNA. In vivo, this drug combination suppressed the growth of tumor xenografts, and this inhibitory effect was significantly decreased in the absence of OGG1. Hence, the present study highlights the roles of base repair enzymes in cell death induction and suggests that the combination of lower doses of ROS-inducing agents with MTH1 inhibitors may be a more selective and safer strategy for cervical cancer chemotherapy.
Sujet(s)
DNA Glycosylases , Enzymes de réparation de l'ADN , Phosphoric monoester hydrolases , Espèces réactives de l'oxygène , Tumeurs du col de l'utérus , Tumeurs du col de l'utérus/traitement médicamenteux , Tumeurs du col de l'utérus/métabolisme , Tumeurs du col de l'utérus/anatomopathologie , Humains , Femelle , Espèces réactives de l'oxygène/métabolisme , Animaux , Phosphoric monoester hydrolases/métabolisme , Phosphoric monoester hydrolases/antagonistes et inhibiteurs , DNA Glycosylases/métabolisme , DNA Glycosylases/antagonistes et inhibiteurs , DNA Glycosylases/génétique , Souris , Enzymes de réparation de l'ADN/métabolisme , Enzymes de réparation de l'ADN/antagonistes et inhibiteurs , Enzymes de réparation de l'ADN/génétique , Guanine/analogues et dérivés , Guanine/pharmacologie , Lignée cellulaire tumorale , Réparation de l'ADN/effets des médicaments et des substances chimiques , Souris nude , Tests d'activité antitumorale sur modèle de xénogreffe , Synergie des médicaments , Cellules HeLa , Stress oxydatif/effets des médicaments et des substances chimiquesRÉSUMÉ
We investigated possible associations between the internal concentrations of POPs and correlations between blood and tumor tissue concentrations in patients who underwent surgery for breast cancer and breast reduction as controls. Genetic variations in CYP1A1, GSTP1, GSTM1, and GSTT1 and hOGG1 were evaluated to determine whether they represent risk factors for breast cancer. Certain POPs have been found to be associated with breast cancer development. GST-P1 polymorphism represented a significant risk for breast cancer with unadjusted OR. However, the GSTT1 null polymorphism represented a significant risk for breast cancer when OR adjusted for age and smoking status. CYP1A1 polymorphism was a significant risk factor for breast cancer, regardless of whether the OR was adjusted. These results suggest that exposure to certain POPs, GSTT1 and CYP1A1 polymorphisms, age, and smoking status are risk factors for breast cancer. In addition, the blood concentrations of some POPs represent surrogates for breast tissue concentrations.