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MicroRNA-30b-mediated regulation of catalase expression in human ARPE-19 cells.

Haque R, Chun E, Howell JC, Sengupta T, Chen D, Kim H - PLoS ONE (2012)

Bottom Line: Here, we demonstrated that a sublethal dose of H(2)O(2) (200 µM) up-regulated the expression of miR-30b, a member of the miR-30 family, which inhibited the expression of endogenous catalase both at the transcript and protein levels.However, antisense (antagomirs) of miR-30b was not only found to suppress the miR-30b mimics-mediated inhibitions, but also to dramatically increase the expression of catalase even under an oxidant environment.We propose that a microRNA antisense approach could enhance cytoprotective mechanisms against oxidative stress by increasing the antioxidant defense system.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Emory University School of Medicine, Atlanta, Georgia, United States of America. rhaque@emory.edu

ABSTRACT

Background: Oxidative injury to retinal pigment epithelium (RPE) and retinal photoreceptors has been linked to a number of retinal diseases, including age-related macular degeneration (AMD). Reactive oxygen species (ROS)-mediated gene expression has been extensively studied at transcriptional levels. Also, the post-transcriptional control of gene expression at the level of translational regulation has been recently reported. However, the microRNA (miRNA/miR)-mediated post-transcriptional regulation in human RPE cells has not been thoroughly looked at. Increasing evidence points to a potential role of miRNAs in diverse physiological processes.

Methodology/principal findings: We demonstrated for the first time in a human retinal pigment epithelial cell line (ARPE-19) that the post-transcriptional control of gene expression via miRNA modulation regulates human catalase, an important and potent component of cell's antioxidant defensive network, which detoxifies hydrogen peroxide (H(2)O(2)) radicals. Exposure to several stress-inducing agents including H(2)O(2) has been reported to alter miRNA expression profile. Here, we demonstrated that a sublethal dose of H(2)O(2) (200 µM) up-regulated the expression of miR-30b, a member of the miR-30 family, which inhibited the expression of endogenous catalase both at the transcript and protein levels. However, antisense (antagomirs) of miR-30b was not only found to suppress the miR-30b mimics-mediated inhibitions, but also to dramatically increase the expression of catalase even under an oxidant environment.

Conclusions/significance: We propose that a microRNA antisense approach could enhance cytoprotective mechanisms against oxidative stress by increasing the antioxidant defense system.

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Related in: MedlinePlus

miR-30 targeting human catalase is sensitive to H2O2.Cultured ARPE-19 cells were treated with vehicle or 200 µM H2O2 for 18 h before harvesting. The expression levels of miR-30 family members were determined by qRT-PCR using snRNA U5 as an internal control. Values are the means ± SEM of relative changes over controls from four samples in each group after normalization to snRNA U5. H2O2 increased the expression levels of miR-30b (p = 0.004) and miR-30d (p = 0.049) miRNAs as compared to the control.
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pone-0042542-g004: miR-30 targeting human catalase is sensitive to H2O2.Cultured ARPE-19 cells were treated with vehicle or 200 µM H2O2 for 18 h before harvesting. The expression levels of miR-30 family members were determined by qRT-PCR using snRNA U5 as an internal control. Values are the means ± SEM of relative changes over controls from four samples in each group after normalization to snRNA U5. H2O2 increased the expression levels of miR-30b (p = 0.004) and miR-30d (p = 0.049) miRNAs as compared to the control.

Mentions: In order to investigate whether the expression of miR-30 family members was influenced by H2O2, ARPE-19 cells were treated with vehicle or 200 µM H2O2 for 18 h. The expression level of miR-30b as determined by qRT-PCR was found to be sensitive to H2O2 (p = 0.004) when compared with the control. The expression of miR-30d was also seen to be up-regulated by H2O2 treatment (p = 0.049). The expression of other members of miR-30 family (miR-30a, miR-30c, and miR-30e) was not observed to be altered by H2O2 treatment, as compared to the control (Figure 4).


MicroRNA-30b-mediated regulation of catalase expression in human ARPE-19 cells.

Haque R, Chun E, Howell JC, Sengupta T, Chen D, Kim H - PLoS ONE (2012)

miR-30 targeting human catalase is sensitive to H2O2.Cultured ARPE-19 cells were treated with vehicle or 200 µM H2O2 for 18 h before harvesting. The expression levels of miR-30 family members were determined by qRT-PCR using snRNA U5 as an internal control. Values are the means ± SEM of relative changes over controls from four samples in each group after normalization to snRNA U5. H2O2 increased the expression levels of miR-30b (p = 0.004) and miR-30d (p = 0.049) miRNAs as compared to the control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3412823&req=5

pone-0042542-g004: miR-30 targeting human catalase is sensitive to H2O2.Cultured ARPE-19 cells were treated with vehicle or 200 µM H2O2 for 18 h before harvesting. The expression levels of miR-30 family members were determined by qRT-PCR using snRNA U5 as an internal control. Values are the means ± SEM of relative changes over controls from four samples in each group after normalization to snRNA U5. H2O2 increased the expression levels of miR-30b (p = 0.004) and miR-30d (p = 0.049) miRNAs as compared to the control.
Mentions: In order to investigate whether the expression of miR-30 family members was influenced by H2O2, ARPE-19 cells were treated with vehicle or 200 µM H2O2 for 18 h. The expression level of miR-30b as determined by qRT-PCR was found to be sensitive to H2O2 (p = 0.004) when compared with the control. The expression of miR-30d was also seen to be up-regulated by H2O2 treatment (p = 0.049). The expression of other members of miR-30 family (miR-30a, miR-30c, and miR-30e) was not observed to be altered by H2O2 treatment, as compared to the control (Figure 4).

Bottom Line: Here, we demonstrated that a sublethal dose of H(2)O(2) (200 µM) up-regulated the expression of miR-30b, a member of the miR-30 family, which inhibited the expression of endogenous catalase both at the transcript and protein levels.However, antisense (antagomirs) of miR-30b was not only found to suppress the miR-30b mimics-mediated inhibitions, but also to dramatically increase the expression of catalase even under an oxidant environment.We propose that a microRNA antisense approach could enhance cytoprotective mechanisms against oxidative stress by increasing the antioxidant defense system.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Emory University School of Medicine, Atlanta, Georgia, United States of America. rhaque@emory.edu

ABSTRACT

Background: Oxidative injury to retinal pigment epithelium (RPE) and retinal photoreceptors has been linked to a number of retinal diseases, including age-related macular degeneration (AMD). Reactive oxygen species (ROS)-mediated gene expression has been extensively studied at transcriptional levels. Also, the post-transcriptional control of gene expression at the level of translational regulation has been recently reported. However, the microRNA (miRNA/miR)-mediated post-transcriptional regulation in human RPE cells has not been thoroughly looked at. Increasing evidence points to a potential role of miRNAs in diverse physiological processes.

Methodology/principal findings: We demonstrated for the first time in a human retinal pigment epithelial cell line (ARPE-19) that the post-transcriptional control of gene expression via miRNA modulation regulates human catalase, an important and potent component of cell's antioxidant defensive network, which detoxifies hydrogen peroxide (H(2)O(2)) radicals. Exposure to several stress-inducing agents including H(2)O(2) has been reported to alter miRNA expression profile. Here, we demonstrated that a sublethal dose of H(2)O(2) (200 µM) up-regulated the expression of miR-30b, a member of the miR-30 family, which inhibited the expression of endogenous catalase both at the transcript and protein levels. However, antisense (antagomirs) of miR-30b was not only found to suppress the miR-30b mimics-mediated inhibitions, but also to dramatically increase the expression of catalase even under an oxidant environment.

Conclusions/significance: We propose that a microRNA antisense approach could enhance cytoprotective mechanisms against oxidative stress by increasing the antioxidant defense system.

Show MeSH
Related in: MedlinePlus