scholarly journals Commutative regulation between endothelial NO synthase and insulin receptor substrate 2 by microRNAs

2018 ◽  
Vol 11 (6) ◽  
pp. 510-521 ◽  
Author(s):  
Xiaoli Sun ◽  
Huizhen Lv ◽  
Peng Zhao ◽  
Jinlong He ◽  
Qinghua Cui ◽  
...  
Keyword(s):  
Cardiovascular Diseases ◽  
Insulin Receptor ◽  
No Synthase ◽  
Luciferase Reporter ◽  
Common Disease ◽  
Enos Gene ◽  
No Production ◽  
Insulin Receptor Substrate ◽  
Endothelial No Synthase ◽  
Mrna Targets

Abstract Endothelial NO synthase (eNOS) expression is regulated by a number of transcriptional and post-transcriptional mechanisms, but the effects of competing endogenous RNAs (ceRNAs) on eNOS mRNA and the underlying mechanisms are still unknown. Our bioinformatic analysis revealed three highly expressed eNOS-targeting miRNAs (miR-15b, miR-16, and miR-30b) in human endothelial cells (ECs). Among the 1103 mRNA targets of these three miRNAs, 15 mRNAs share a common disease association with eNOS. Gene expression and correlation analysis in patients with cardiovascular diseases identified insulin receptor substrate 2 (IRS2) as the most correlated eNOS-ceRNA. The expression levels of eNOS and IRS2 were coincidentally increased by application of laminar shear but reduced with eNOS or IRS2 siRNA transfection in human ECs, which was impeded by Dicer siRNA treatment. Moreover, luciferase reporter assay showed that these three miRNAs directly target the 3′UTR of eNOS and IRS2. Overexpression of these three miRNAs decreased, whereas inhibition of them increased, both mRNA and protein levels of eNOS and IRS2. Functionally, silencing eNOS suppressed the Akt signal pathway, while IRS2 knockdown reduced NO production in ECs. Thus, we identified eNOS and IRS2 as ceRNAs and revealed a novel mechanism explaining the coincidence of metabolic and cardiovascular diseases.

17β-Estradiol decreases hypoxic induction of erythropoietin gene expression

2002 ◽  
Vol 283 (2) ◽  
pp. R496-R504 ◽  
Author(s):  
Harshini Mukundan ◽  
Thomas C. Resta ◽  
Nancy L. Kanagy
Keyword(s):  
Gene Expression ◽  
Nitric Oxide ◽  
Oxygen Delivery ◽  
Chronic Hypoxia ◽  
No Synthase ◽  
No Production ◽  
Enos Expression ◽  
Hypoxic Induction ◽  
Endothelial No Synthase ◽  
17Β Estradiol

Exposure to chronic hypoxia induces erythropoietin (EPO) production to facilitate oxygen delivery to hypoxic tissues. Previous studies from our laboratory found that ovariectomy (OVX) exacerbates the polycythemic response to hypoxia and treatment with 17β-estradiol (E2-β) inhibits this effect. We hypothesized that E2-β decreases EPO gene expression during hypoxia. Because E2-β can induce nitric oxide (NO) production and NO can attenuate EPO synthesis, we further hypothesized that E2-β inhibition of EPO gene expression is mediated by NO. These hypotheses were tested in OVX catheterized rats treated with E2-β (20 μg/day) or vehicle for 14 days and exposed to 8 or 12 h of hypoxia (12% O2) or normoxia. We found that E2-β treatment significantly decreased EPO synthesis and gene expression during hypoxia. E2-β treatment did not induce endothelial NO synthase (eNOS) expression in the kidney but potentiated hypoxia-induced increases in plasma nitrates. We conclude that E2-β decreases hypoxic induction of EPO. However, this effect does not appear to be related to changes in renal eNOS expression.

Diabetic HDL-associated myristic acid inhibits acetylcholine-induced nitric oxide generation by preventing the association of endothelial nitric oxide synthase with calmodulin

2008 ◽  
Vol 294 (1) ◽  
pp. C295-C305 ◽  
Author(s):  
James White ◽  
Theresa Guerin ◽  
Hollie Swanson ◽  
Steven Post ◽  
Haining Zhu ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Myristic Acid ◽  
Acetylcholine Receptors ◽  
Diabetic Mouse ◽  
No Synthase ◽  
Diabetic Patients ◽  
No Production ◽  
Dependent Manner ◽  
Endothelial No Synthase ◽  
Stimulation Of

In the current study, we examined whether diabetes affected the ability of HDL to stimulate nitric oxide (NO) production. Using HDL isolated from both diabetic humans and diabetic mouse models, we found that female HDL no longer induced NO synthesis, despite containing equivalent amounts of estrogen as nondiabetic controls. Furthermore, HDL isolated from diabetic females and males prevented acetylcholine-induced stimulation of NO generation. Analyses of both the human and mouse diabetic HDL particles showed that the HDLs contained increased levels of myristic acid. To determine whether myristic acid associated with HDL particles was responsible for the decrease in NO generation, myristic acid was added to HDL isolated from nondiabetic humans and mice. Myristic acid-associated HDL inhibited the generation of NO in a dose-dependent manner. Importantly, diabetic HDL did not alter the levels of endothelial NO synthase or acetylcholine receptors associated with the cells. Surprisingly, diabetic HDL inhibited ionomycin-induced stimulation of NO production without affecting ionomycin-induced increases in intracellular calcium. Further analysis indicated that diabetic HDL prevented calmodulin from interacting with endothelial NO synthase (eNOS) but did not affect the activation of calmodulin kinase or calcium-independent mechanisms for stimulating eNOS. These studies are the first to show that a specific fatty acid associated with HDL inhibits the stimulation of NO generation. These findings have important implications regarding cardiovascular disease in diabetic patients.

scholarly journals Apolipoprotein E Enhances Endothelial-NO Production by Modulating Caveolin 1 Interaction With Endothelial NO Synthase

Hypertension ◽  
2012 ◽  
Vol 60 (4) ◽  
pp. 1040-1046 ◽  
Author(s):  
Lili Yue ◽  
Jing-Tan Bian ◽  
Ivana Grizelj ◽  
Ana Cavka ◽  
Shane A. Phillips ◽  
...  
Keyword(s):  
Apolipoprotein E ◽  
No Synthase ◽  
No Production ◽  
Caveolin 1 ◽  
Endothelial No Synthase

scholarly journals Role of MicroRNA 30a Targeting Insulin Receptor Substrate 2 in Colorectal Tumorigenesis

2015 ◽  
Vol 35 (6) ◽  
pp. 988-1000 ◽  
Author(s):  
Qian Zhang ◽  
Qingchao Tang ◽  
Dandan Qin ◽  
Lei Yu ◽  
Rui Huang ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Insulin Receptor ◽  
Cell Lines ◽  
Cancer Cell ◽  
Colon Cancer Cell ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Insulin Receptor Substrate ◽  
Human Colon Cancer

MicroRNAs (miRNAs) are dysregulated in many types of malignant diseases, including colorectal cancer. miRNA 30a (miR-30a) is a member of the miR-30 family and has been implicated in many types of cancers. In this study, we determined the expression of miR-30a in human colon cancer tissues and cell lines. miR-30a was found to be significantly downregulated in both the tissues and cell lines. Furthermore, overexpression of miR-30a inhibited, while silencing of miR-30a promoted, cell proliferation, migration, and invasion in vitro . Consistently, stable overexpression of miR-30a suppressed the growth of colon cancer cell xenografts in vivo . Moreover, bioinformatic algorithms and luciferase reporter assays revealed that insulin receptor substrate 2 (IRS2) is a direct target of miR-30a. Further functional studies suggested that repression of IRS2 by miR-30a partially mediated the tumor suppressor effect of miR-30a. In addition, miR-30a inhibited constitutive phosphorylation of Akt by targeting IRS2. Additionally, clinicopathological analysis indicated that miR-30a has an inverse correlation with the staging in patients with colon cancer. Taken together, our study provides the first evidence that miR-30a suppressed colon cancer cell growth through inhibition of IRS2. Thus, miR-30a might serve as a promising therapeutic strategy for colon cancer treatment.

The significance of −786T>Cpolymorphism of endothelial NO synthase (eNOS) gene in severe preeclampsia

2010 ◽  
Vol 24 (3) ◽  
pp. 432-436 ◽  
Author(s):  
Agnieszka Seremak-Mrozikiewicz ◽  
Krzysztof Drews ◽  
Magdalena Barlik ◽  
Piotr Sieroszewski ◽  
Edmund Grześkowiak ◽  
...  
Keyword(s):  
No Synthase ◽  
Severe Preeclampsia ◽  
Enos Gene ◽  
Endothelial No Synthase

Palmitic Acid Regulates miRNA-3148 via Insulin Receptor Substrate-1 and is Involved in Insulin Resistance

2021 ◽  
Vol 11 (4) ◽  
pp. 767-771
Author(s):  
Xiaoxi Xiang ◽  
Changwei Zhang ◽  
Daying Long
Keyword(s):  
Insulin Resistance ◽  
Palmitic Acid ◽  
Insulin Receptor ◽  
Hepg2 Cells ◽  
Luciferase Reporter ◽  
Insulin Receptor Substrate ◽  
Insulin Receptor Substrate 1 ◽  
Over Expression ◽  
Reporter Assays ◽  
The Impact

Previous studies reported that saturated fatty acid palmitic acid (PA) is closely related to insulin resistance. miR-3148 regulates insulin receptor substrate-1 (IRS1) predicted by MiRDB analysis. However, whether PA regulates IRS1 via miR-3148 remains to be elucidated. Therefore, in this work, we assessed whether PA regulates miRNA-3148 via IRS1 in insulin resistance. We cultured HepG2 cells in vitro and classified them into control group (NC group), miR-3148 Mimics group, and miR-3148 Mimics+ pFBD-IRS1 group. We used qRT-PCR to detect miR-3148 and IRS1 mRNA; used Dual-Luciferase Reporter Assays to detect miR-3148 with 3′-UTR region of IRS1 mRNA; and utilized Western blot (WB) to detect IRS1, p-AKT, AKT and Tubulin. Our results showed that PA could increase miR-3148 and decrease IRS1 which is a target protein of miR-3148, as shown by Dual-Luciferase Reporter assays. miR-3148 significantly inhibited the impact of insulin on p-AKT level (P < 0.01) and over-expression of IRS1 by pFBD-IRS1 can partially alleviate the inhibitory effect of miR-3148 mimics on p-AKT. In HepG2 cells, PA regulates miR-3148. Via targeting IRS1 mRNA, miR-3148 impairs insulin signaling pathway, leading to insulin resistance. Over-expression of IRS1 by pFBD-IRS1 alleviates miR-3148-induced insulin resistance.

P6603The predictive role of T786C single nucleotide polymorphism in endothelial no-synthase gene in late left ventricular remodeling after ST-segment elevation myocardial infarction

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
O Petyunina ◽  
M P Kopytsya ◽  
A E Berezin ◽  
A A Berezin
Keyword(s):  
Myocardial Infarction ◽  
Single Nucleotide Polymorphism ◽  
Cardiac Remodeling ◽  
Independent Predictor ◽  
No Synthase ◽  
Enos Gene ◽  
Ethical Standards ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Endothelial No Synthase

Abstract Introduction Endothelial NO-synthase (eNOS) is constitutive enzyme, which is expressed in mature endothelial cells and promotes direct vascular dilatation. Single nucleotide polymorphism (SNP) of T786C in eNOS gene may influence on adverse cardiac remodeling after ST-elevation myocardial infarction (STEMI). Purpose To investigate possible associations between SNP T786C in eNOS gene and adverse cardiac remodeling after STEMI Methods 177 acute STEMI patients treated with primary and facilitate percutaneous coronary intervention that were admitted to intensive care unit of a Therapy National Institute were enrolled in the study. Anthropometry, cardiovascular risk assay, coronary angiography, echocardiography and biomarkers' measure were performed at baseline. The DNA extraction was performed with a commercial kit using real-time polymerase chain reaction PCR. All procedures performed in the study involving human participants were in accordance with the ethical standards and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards and approved by the local ethics committee (Protocol No. 8, 29.08.2016). Written informed consent was obtained from each patient. Results There were correlations between 786CC polymorphism in eNOs gene and adverse cardiac remodeling (r=0.48; p=0.001), LDL cholesterol (r=0.32; p=0.012), type 2 diabetes mellitus (r=0.30; p=0.042), diastolic BP (r=−0.26; p=0.048), unstable angina prior to STEMI (r=0.25; p=0.047) and total quantity of complicated STEMI (r=0.23; p=0.042). Additionally, there were not significant relations between 786CC polymorphism in eNOs gene and multiple coronary vessel injury, STEMI localization, levels of circulating biomarkers of myocardial injury, and amount of damaged coronary arteries. Using univariate and multivariate regressive logistic analysis we found that 786CC genotype of eNOS was independent predictor for late adverse LV remodeling (β-coefficient = 1.57342; odds ratio = 4.8231; 95% confidence interval = 1.5349–15.1552; p=0.0071). Conclusions The polymorphism 786CC in eNOs gene was found as an independent predictor for late adverse cardiac remodeling after STEMI. Acknowledgement/Funding None

scholarly journals Knockdown of long noncoding RNA PVT1 suppresses cell proliferation and invasion of colorectal cancer via upregulation of microRNA-214-3p

2019 ◽  
Vol 317 (2) ◽  
pp. G222-G232 ◽  
Author(s):  
An-Quan Shang ◽  
Wei-Wei Wang ◽  
Yi-Bao Yang ◽  
Chen-Zheng Gu ◽  
Ping Ji ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Proliferation ◽  
Insulin Receptor ◽  
Luciferase Reporter ◽  
Insulin Receptor Substrate ◽  
Insulin Receptor Substrate 1 ◽  
Gene Assay ◽  
Proliferation And Invasion ◽  
Variant Translocation ◽  
Competitive Endogenous Rna

Long noncoding RNAs (lncRNAs) have been reported to be involved in the occurrence and tumorigenesis of numerous malignant cancers. Microarray expression profiles were used to screen colorectal cancer (CRC)-related differentially expressed genes and lncRNAs, which revealed that insulin receptor substrate 1 (IRS1) and lncRNA plasmacytoma variant translocation 1 (PVT1) were highly expressed in CRC. This study aimed to investigate the regulatory role of lncRNA PVT1 in CRC. Subcellular localization detected by fluorescence in situ hybridization identified that lncRNA PVT1 was primarily located in the cytoplasm. The interaction between lncRNA PVT1 and microRNA-214-3p (miR-214-3p) and IRS1 was predicted using the RNA22 website. Next the dual luciferase reporter gene assay, RNA pull-down, and RNA immunoprecipitation assays verified lncRNA PVT1 to be a competitive endogenous RNA (ceRNA) against miR-214-3p, and IRS1 was found to be a target of miR-214-3p. The expression pattern of lncRNA PVT1, miR-214-3p, IRS1, phosphoinositide 3-kinase (PI3K), and Akt was characterized in response to lncRNA PVT1 silencing or miR-214-3p upregulation. Meanwhile, their regulatory effects on cell proliferation, invasion, and apoptosis were detected in CRC cells. With increased levels of miR-214-3p and decreased levels of lncRNA PVT1 in CRC cells, the expression of phosphatidylinositol 3-kinase, putative (PI3K) and Akt was reduced, and consequently, the cell apoptosis was stimulated and cell proliferation and invasion were suppressed. All in all, lncRNA PVT1 competitively binds to miR-214-3p to upregulate the expression of IRS1 thus activating the PI3K/Akt signaling pathway, thus accelerating CRC progression. This study suggests that lncRNA PVT1 might be a potential target of therapeutic strategies for CRC treatment. NEW & NOTEWORTHY This study mainly suggests that long noncoding (lnc)RNA plasmacytoma variant translocation 1 (PVT1) is a downregulated lncRNA in colorectal cancer (CRC), accelerating CRC progression. Strikingly, lncRNA PVT1 acts as a competitive endogenous RNA against microRNA (miR)-214-3p, whereas miR-214-3p targets insulin receptor substrate 1, which draws a comprehensive picture of the potential molecular mechanisms of lncRNA PVT1 in CRC.

scholarly journals Cell Cycle-Regulated Inactivation of Endothelial NO Synthase through NOSIP-Dependent Targeting to the Cytoskeleton

2005 ◽  
Vol 25 (18) ◽  
pp. 8251-8258 ◽  
Author(s):  
Michael Schleicher ◽  
Fredrik Brundin ◽  
Steffen Gross ◽  
Werner Müller-Esterl ◽  
Stefanie Oess
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Nuclear Localization ◽  
Nuclear Export ◽  
Vascular Function ◽  
No Synthase ◽  
Nuclear Localization Sequence ◽  
No Production ◽  
Endothelial No Synthase ◽  
Importin Α

ABSTRACT Nitric oxide (NO) plays a key role in vascular function, cell proliferation, and apoptosis. Proper subcellular localization of endothelial NO synthase (eNOS) is crucial for its activity; however, the role of eNOS trafficking for NO biosynthesis remains to be defined. Overexpression of NOS-interacting protein (NOSIP) induces translocation of eNOS from the plasma membrane to intracellular compartments, thereby impairing NO production. Here we report that endogenous NOSIP reduces the enzymatic capacity of eNOS, specifically in the G2 phase of the cell cycle by targeting eNOS to the actin cytoskeleton. This regulation is critically dependent on the nucleocytoplasmic shuttling of NOSIP and its cytoplasmic accumulation in the G2 phase. The predominant nuclear localization of NOSIP depends on a bipartite nuclear localization sequence (NLS) mediating interaction with importin α. Mutational destruction of the NLS abolishes nuclear import and interaction with importin α. Nuclear export is insensitive to leptomycin B and hence different from the CRM1-dependent default mechanism. Inhibition of NOSIP expression by RNA interference completely abolishes G2-specific cytoskeletal association and inhibition of eNOS. These findings describe a novel cell cycle-dependent modulation of endogenous NO levels that are critical to the cell cycle-related actions of NO such as apoptosis or cell proliferation.

scholarly journals Neuronal insulin receptor substrate 2 (IRS2) expression is regulated by ZBP89 and SP1 binding to the IRS2 promoter

2009 ◽  
Vol 204 (2) ◽  
pp. 199-208 ◽  
Author(s):  
Michael Udelhoven ◽  
Mareike Pasieka ◽  
Uschi Leeser ◽  
Wilhelm Krone ◽  
Markus Schubert
Keyword(s):  
Insulin Receptor ◽  
Binding Sites ◽  
Luciferase Reporter ◽  
Serum Starvation ◽  
Insulin Receptor Substrate ◽  
Dependent Manner ◽  
Reporter Assays ◽  
Phosphoinositide 3 Kinase ◽  
Translational Start ◽  
Specificity Protein

Since neuronal insulin receptor substrate 2 (IRS2)-mediated signals coordinate key processes in rodent physiology such as food intake, fertility, longevity, and aging-related behavior, we analyzed the mechanisms of neuronal IRS2 expression in neuroblastoma (SHSY5Y) and hypothalamic (GT1-7) cell lines. Using dual luciferase reporter assays and IRS2 promoter deletion constructs, we identified a regulatory cassette within the IRS2 promoter between −779 and −679 bp from the translational start which is responsible for ∼50% of neuronal IRS2 promoter activity. Chromatin immunoprecipitation assays and electromobility shift assay revealed four overlapping ZBP89/specificity protein 1 (SP1) binding sites which alternatively bind to ZBP89 (ZNF148 as listed in the HUGO Database) or SP1. Activation of this cassette is inhibited by phosphoinositide-3-kinase (PI3K) via increased ZBP89 binding to the promoter. Serum starvation caused increased SP1 binding at one specific SP1 site and decreased binding to another, proving a regulatory interaction between the different binding sites within this promoter cassette to tightly control IRS2 expression. Mutants containing all the possible combinations of one, two, three, or all the four SP1 binding sites of the IRS2 promoter revealed that SP1 binding to one particular site is most important for promoter activation. Stable downregulation of ZBP89 using siRNA substantially increased IRS2 mRNA and protein expression. Thus, alternative binding of ZBP89 or SP1 to the described region in the IRS2 promoter regulates neuronal IRS2 expression in a PI3K-dependent manner.

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