5-aza-2′-deoxycytidine Inhibits the Proliferation of Lung Fibroblasts in Neonatal Rats Exposed to Hyperoxia

Cao, Meiling; Wu, Hongmin; Yu, Huimei; Xue, Xindong

doi:10.1016/j.pedneo.2015.11.009

Cited by 17 publications

(16 citation statements)

References 18 publications

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“…The concentrations of NaF and the time of 72 hours were chosen based on data obtained by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2.5‐iphenyltetrazolium bromide (MTT) assays. To determine if the changes of gene expression and cell cycle were affected by aberrant methylation, cells were treated with NaF together with 5‐AZA‐dC (Sigma, Missouri) at 5, 10, or 20 μmol·L −1 , for 72 hours …”

Section: Materirals and Methodsmentioning

confidence: 99%

“…Furthermore, DNA methylation that silences gene expression is a reversible process. 5‐aza‐2‐deoxycytidine (5‐AZA‐dC), used for reversing DNA methylation, reactivates suppressor genes silenced by promoter methylation . 5‐AZA‐dC is utilized for the treatment of myelodysplastic syndrome and has been tested in clinical trials for treatment of acute and chronic myelogenous leukemia …”

Section: Introductionmentioning

confidence: 99%

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Aberrant methylation‐induced dysfunction of p16 is associated with osteoblast activation caused by fluoride

Qiu

et al. 2018

Environmental Toxicology

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Chronic exposure to fluoride continues to be a public health problem worldwide, affecting thousands of people. Fluoride can cause abnormal proliferation and activation of osteoblast and osteoclast, leading to skeletal fluorosis that can cause pain and harm to joints and bones and even lead to permanent disability. Nevertheless, there is no recognized mechanism to explain the bone lesions of fluorosis. In this work, we performed a population study and in vitro experiments to investigate the pathogenic mechanism of skeletal fluorosis in relation to methylation of the promoter of p16. The protein coded by the p16 gene inhibits cdk (cyclin-dependent kinase) 4/cdk6-mediated phosphorylation4 of retinoblastoma gene product and induces cell cycle arrest.The results showed that hypermethylation of p16 and reduced gene expression was evident in peripheral blood mononuclear cells of patients with fluorosis and correlated with the level of fluoride exposure. Studies with cell cultures of osteoblasts revealed in response to sodium fluoride (NaF) treatment, there was an induction of p16 hypermethylation and decreased expression, leading to increased cell proliferation, a longer S-phase of the cell cycle, and development of skeletal fluorosis. Further, the methylation inhibitor, 5-aza-2-deoxycytidine, reversed the p16 hypermethylation and expression in response to NaF. These results reveal a regulatory role of p16 gene methylation on osteoblasts activation during the development of skeletal fluorosis. K E Y W O R D S coal-burning fluorosis, DNA methylation, osteoblast activation, p16 gene, skeletal fluorosis

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Section: Materirals and Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Aberrant methylation‐induced dysfunction of p16 is associated with osteoblast activation caused by fluoride

Qiu

et al. 2018

Environmental Toxicology

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“…Cells were cultured in a 96-well plate, following incubation with atorvastatin at different concentrations, namely 0.5, 1.0, 1.5, and 2.0 lM, at 37°C with air containing 5% CO 2 for about 24 h. Then, CCK8 (Solarbio, Beijing, China) reagent was added and the attenuance was measured at 570 nm and the percentage cell viability was calculated as described previously [56]. Cells were cultured in a 96-well plate, following incubation with atorvastatin at different concentrations, namely 0.5, 1.0, 1.5, and 2.0 lM, at 37°C with air containing 5% CO 2 for about 24 h. Then, CCK8 (Solarbio, Beijing, China) reagent was added and the attenuance was measured at 570 nm and the percentage cell viability was calculated as described previously [56].…”

Section: Cck8 Assaymentioning

confidence: 99%

“…Cells with different treatments were plated in a 96-well plate and the cell proliferation detected with Cell Counting Kit-8 (Beyotime Institute of Biotechnology, Shanghai, China) according to the manufacturer's instructions. Cells were cultured in a 96-well plate, following incubation with atorvastatin at different concentrations, namely 0.5, 1.0, 1.5, and 2.0 lM, at 37°C with air containing 5% CO 2 for about 24 h. Then, CCK8 (Solarbio, Beijing, China) reagent was added and the attenuance was measured at 570 nm and the percentage cell viability was calculated as described previously [56].…”

Section: Cck8 Assaymentioning

confidence: 99%

Atorvastatin treatment modulates p16 promoter methylation to regulate p16 expression

et al. 2017

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Intimal hyperplasia, the key event of arterial restenosis, is a result of cell proliferation and cell migration. Atorvastatin exerts an inhibitory effect on cell proliferation and migration, but the mechanism remains largely unknown. p16, as a well-known tumor suppressor, was also reported to suppress cell growth and migration, but with an unclear mechanism. In this study, we demonstrated that atorvastatin represses cell proliferation and migration in vascular smooth muscle cells (VSMCs) and that this process is mediated by p16. Furthermore, we found that DNA methylation in the p16 promoter was reduced and p16 expression was restored in VSMCs treated with 5-aza-2'-deoxycytidine or atorvastatin. However, the effect was absent when DNA methyltransferase 1 (DNMT1) was knocked down with RNA interference. These observations demonstrated that atorvastatin regulates p16 expression via DNMT1-induced DNA methylation in the p16 promoter. In addition, we found that the mitogen-activated protein kinase (MAPK) pathway was involved in the regulation of p16 by DNMT1, and MAPK inhibitors partially released the effects of atorvastatin on p16 and DNMT1. Finally, we illustrated that atorvastatin inhibits neointima formation and modulates p16 expression in balloon catheter-injured rat carotid artery. Taken together, we demonstrated that atorvastatin inhibits neointima formation through inducing p16 expression by affecting DNA methylation in the p16 promoter region.

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“…It combines with DNA during DNA replication, forms covalent complexes with DNA methyltransferase 1 (DNMT1), inhibits the methyl transfer activity of the enzyme, produces low-methyl annihilator chains, and reduces hypermethylation of the gene promoter region to recover gene activity (11,17,18). Our previous study also indicated that 5-aza-CdR inhibits the growth of the LFs in hyperoxia-induced neonatal BPD rats in vitro by demethylating the P16 gene (19). Therefore, the aim of this study was to evaluate the possible preventive effect of 5-aza-CdR in a neonatal rat model of hyperoxia lung fibrosis and to elucidate the relationship between P16 gene expression and lung fibrosis induced by hyperoxia.…”

mentioning

confidence: 99%

5-aza-2′-deoxycytidine, a DNA methylation inhibitor, attenuates hyperoxia-induced lung fibrosis via re-expression of P16 in neonatal rats

Cao

Han

2017

Pediatr Res

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BackgroundP16 methylation plays an important role in the pathogenesis of hyperoxia-induced lung fibrosis. 5-aza-2'-deoxycytidine (5-aza-CdR) is a major methyltransferase-specific inhibitor. In this study, the effects of 5-aza-CdR on a hyperoxia-induced lung fibrosis in neonatal rats were investigated.MethodsRat pups were exposed to 85% O for 21 days of and received intraperitoneal injections of 5-aza-CdR or normal saline (NS) once every other day. Survival rates and lung coefficients were calculated. Hematoxylin-eosin staining was performed to analyze the degree of lung fibrosis. Collagen content and TGF-β1 levels were determined. A methylation-specific polymerase chain reaction and western blotting were performed to determine P16 methylation status and P16, cyclin D1, and E2F1 protein expression.Results5-aza-CdR treatment during hyperoxia significantly improved the survival rate and weight gain, while it decreases the degree of lung fibrosis and levels of hydroxyproline and TGF-β1. Hyperoxia induced abnormal P16 methylation and 5-aza-CdR effectively reversed the hypermethylation of P16. Expression of the P16 protein in lung tissues was enhanced, while cyclin D1 and E2F1 protein were reduced by 5-aza-CdR treatment during hyperoxia.ConclusionThese data show that 5-aza-CdR attenuated lung fibrosis in neonatal rats exposed to hyperoxia by lowering hydroxyproline and TGF-β1 expression and via re-expression of P16 in neonatal rats.

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5-aza-2′-deoxycytidine Inhibits the Proliferation of Lung Fibroblasts in Neonatal Rats Exposed to Hyperoxia

Abstract: 5-aza-CdR inhibits the growth of the LFs in hyperoxia-induced neonatal BPD rats in vitro by demethylating the P16 gene.

Cited by 17 publications

References 18 publications

Aberrant methylation‐induced dysfunction of p16 is associated with osteoblast activation caused by fluoride

Aberrant methylation‐induced dysfunction of p16 is associated with osteoblast activation caused by fluoride

Atorvastatin treatment modulates p16 promoter methylation to regulate p16 expression

5-aza-2′-deoxycytidine, a DNA methylation inhibitor, attenuates hyperoxia-induced lung fibrosis via re-expression of P16 in neonatal rats

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