MicroRNA-22 regulates smooth muscle cell differentiation from stem cells by targeting methyl CpG binding protein 2

Zhao, Hanqing; Wen, G.; Huang, Yuan; Yu, Xinyan; Chen, Qishan; Afzal, Tayyab Adeel; Luong, Le Anh; Zhu, Jiang; Zhang, Li; Xiao, Qingzhong

doi:10.1016/j.atherosclerosis.2014.10.080

Cited by 13 publications

(17 citation statements)

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“…miRNA Mimics and Inhibitor Transfection miRNA mimics and inhibitor transfection were conducted as described previously with some modifications. 8,23,29,30 Briefly, either miRNA mimics or inhibitors and miRNA-negative controls (25 nmol/L, final concentration) were transfected into VSMCs using TransIT-X2 transfection reagent (Geneflow Ltd), according to the manufacturer's instructions. VSMCs (1.5910 5 per well) were seeded into 6-well plate 24 hours prior to transfection.…”

Section: Vsmc Culture and Treatmentsmentioning

confidence: 99%

“…Luciferase assays for various gene 3 0 UTR reporters were conducted as described in our previous studies. 8,23,29,30,32 Briefly, VSMCs were cotransfected with an individual reporter gene (pmiR-Luc-NCKAP1-WT, pmiR-Luc-NCKAP1-BS1 mut , pmiR-Luc-NCKAP1-BS2 mut , pmiR-Luc-NCKAP1-BS3 mut , pmiR-Luc-Notch1-BS1/2/3 mut , pmiR-Luc-KLF14, pmiR-Luc-SMYD5, or pmiR-Luc-Notch1; 0.15 lg/2.5910 4 cells) and control or miR-214 (or miR-34a) mimics (25 nmol/L) using TransIT-X2 transfection reagent (Geneflow Ltd), according to the manufacturer's instructions. pmiR-Luc-b-gal (0.20 lg/2.5910 4 cells) or Renilla plasmid (15 ng/well) was included in all transfection assays as an internal control.…”

Section: Transient Transfection and Luciferase Assaymentioning

confidence: 99%

“…Real-time quantitative PCR (RT-qPCR) was performed as described previously. 8,23,29,30,36 Briefly, total RNAs containing small RNAs (miRNAs) were extracted from cells using the mirVana Protein and RNA Isolation System Kit (Thermo Fisher Scientific Inc) or TRI reagent (Sigma-Aldrich), according to the manufacturer's instructions, and subjected to DNase I (Sigma-Aldrich) digestion to remove potential DNA contamination. Reverse transcription for long RNA was performed using an Improm-II RT kit (Promega) with RNase inhibitor (Promega) and Random primers (Promega).…”

Section: Reverse Transcriptase-quantitative Pcr For Mrna and Mirnasmentioning

confidence: 99%

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NCK Associated Protein 1 Modulated by miRNA‐214 Determines Vascular Smooth Muscle Cell Migration, Proliferation, and Neointima Hyperplasia

Afzal

Luong

Chen

et al. 2016

JAHA

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BackgroundMicroRNA miR‐214 has been implicated in many biological cellular functions, but the impact of miR‐214 and its target genes on vascular smooth muscle cell (VSMC) proliferation, migration, and neointima smooth muscle cell hyperplasia is unknown.Methods and ResultsExpression of miR‐214 was closely regulated by different pathogenic stimuli in VSMCs through a transcriptional mechanism and decreased in response to vascular injury. Overexpression of miR‐214 in serum‐starved VSMCs significantly decreased VSMC proliferation and migration, whereas knockdown of miR‐214 dramatically increased VSMC proliferation and migration. Gene and protein biochemical assays, including proteomic analyses, showed that NCK associated protein 1 (NCKAP1)—a major component of the WAVE complex that regulates lamellipodia formation and cell motility—was negatively regulated by miR‐214 in VSMCs. Luciferase assays showed that miR‐214 substantially repressed wild‐type but not the miR‐214 binding site mutated version of NCKAP1 3′ untranslated region luciferase activity in VSMCs. This result confirmed that NCKAP1 is the functional target of miR‐214 in VSMCs. NCKAP1 knockdown in VSMCs recapitulates the inhibitory effects of miR‐214 overexpression on actin polymerization, cell migration, and proliferation. Data from cotransfection experiments also revealed that inhibition of NCKAP1 is required for miR‐214–mediated lamellipodia formation, cell motility, and growth. Importantly, locally enforced expression of miR‐214 in the injured vessels significantly reduced NCKAP1 expression levels, inhibited VSMC proliferation, and prevented neointima smooth muscle cell hyperplasia after injury.ConclusionsWe uncovered an important role of miR‐214 and its target gene NCKAP1 in modulating VSMC functions and neointima hyperplasia. Our findings suggest that miR‐214 represents a potential therapeutic target for vascular diseases.

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Section: Vsmc Culture and Treatmentsmentioning

confidence: 99%

Section: Transient Transfection and Luciferase Assaymentioning

confidence: 99%

Section: Reverse Transcriptase-quantitative Pcr For Mrna and Mirnasmentioning

confidence: 99%

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NCK Associated Protein 1 Modulated by miRNA‐214 Determines Vascular Smooth Muscle Cell Migration, Proliferation, and Neointima Hyperplasia

Afzal

Luong

Chen

et al. 2016

JAHA

Self Cite

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“…Therefore, characterizing the regulation of vascular SMC differentiation is essential for both the prevention and treatment of these diseases. Although recent studies have improved the current understanding of SMC differentiation and cardiovascular system development [2][3][4][5][6][7][8][9][10][11][12], the molecular mechanisms governing SMC differentiation from pluripotent stem cells remain largely unknown.…”

Section: Introductionmentioning

confidence: 99%

Glycoprotein M6B Interacts with TβRI to Activate TGF-β-Smad2/3 Signaling and Promote Smooth Muscle Cell Differentiation

et al. 2018

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Smooth muscle cells (SMCs), which form the walls of blood vessels, play an important role in vascular development and the pathogenic process of vascular remodeling. However, the molecular mechanisms governing SMC differentiation remain poorly understood. Glycoprotein M6B (GPM6B) is a four‐transmembrane protein that belongs to the proteolipid protein family and is widely expressed in neurons, oligodendrocytes, and astrocytes. Previous studies have revealed that GPM6B plays a role in neuronal differentiation, myelination, and osteoblast differentiation. In the present study, we found that the GPM6B gene and protein expression levels were significantly upregulated during transforming growth factor‐β1 (TGF‐β1)‐induced SMC differentiation. The knockdown of GPM6B resulted in the downregulation of SMC‐specific marker expression and repressed the activation of Smad2/3 signaling. Moreover, GPM6B regulates SMC Differentiation by Controlling TGF‐β‐Smad2/3 Signaling. Furthermore, we demonstrated that similar to p‐Smad2/3, GPM6B was profoundly expressed and coexpressed with SMC differentiation markers in embryonic SMCs. Moreover, GPM6B can regulate the tightness between TβRI, TβRII, or Smad2/3 by directly binding to TβRI to activate Smad2/3 signaling during SMC differentiation, and activation of TGF‐β‐Smad2/3 signaling also facilitate the expression of GPM6B. Taken together, these findings demonstrate that GPM6B plays a crucial role in SMC differentiation and regulates SMC differentiation through the activation of TGF‐β‐Smad2/3 signaling via direct interactions with TβRI. This finding indicates that GPM6B is a potential target for deriving SMCs from stem cells in cardiovascular regenerative medicine. Stem Cells 2018 Stem Cells 2019;37:190–201

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“…2–4 The miRs can stimulate NRVM hypertrophy, 4, 5 protect NRVMs from apoptosis by downregulating NCX, 6 reduce cardiac injury, and stimulate adaptive vasculogenesis. 7–10 Studies link reverse remodeling after MI to miR-132 induction and MeCP2 repression; 7 to an adaptive response in the bladder by miR-132/212 induction and MeCP2 repression; 11 and to liver fibrosis by miR 132 repression, MeCP2 induction, and PPAR gamma downregulation. 12 Downregulation of MeCP2 by siRNA reduces apoptosis after ischemia, 13 and MeCP2 overexpression in heart can be embryonic lethal, 14 in agreement with the current study.…”

Section: Mirs-132/212 and Mecp2: Background And Supporting Datamentioning

confidence: 99%

A New Pathway for Sympathetic Cardioprotection in Heart Failure

Simpson

2015

Circulation Research

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MicroRNA-22 regulates smooth muscle cell differentiation from stem cells by targeting methyl CpG binding protein 2

Cited by 13 publications

References 9 publications

NCK Associated Protein 1 Modulated by miRNA‐214 Determines Vascular Smooth Muscle Cell Migration, Proliferation, and Neointima Hyperplasia

NCK Associated Protein 1 Modulated by miRNA‐214 Determines Vascular Smooth Muscle Cell Migration, Proliferation, and Neointima Hyperplasia

Glycoprotein M6B Interacts with TβRI to Activate TGF-β-Smad2/3 Signaling and Promote Smooth Muscle Cell Differentiation

A New Pathway for Sympathetic Cardioprotection in Heart Failure

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