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The intricate immunomodulatory mechanism underlying the onset and progression of chronic rhinosinusitis with nasal polyps (CRSwNP) in chronic sinusitis with nasal polyps remains poorly understood, particularly regarding the regulatory role of dendritic cells (DCs). MicroRNA has been identified as a modulator of DC function. Primary cells have a short time in vitro and their biological characteristics have not changed greatly which can better reflect the growth state of cells in the body, so as to obtain data closer to the physiological function in the body. Our research group has been engaged in the culture of human nasal epithelial progenitor cells (hNEPCs) for a long time, and in previous investigations, we observed downregulation of microRNA‐124 (miR‐124) in patients with chronic sinusitis and nasal polyps. Thus, our current study aims to elucidate the regulatory mechanism of DCs in chronic sinusitis with polyps by examining the impact of miR‐124 on DCs in this context. To accomplish this, we took the nasal mucosa of CRSwNP patients and normal nasal mucosa of non‐rhinitis patients for primary culture. We isolated exosomes from these cells for co‐culture experiments with DCs. Flow cytometry analysis was employed to assess the maturation phenotypes (MHC II, CD80 and CD86), cell cycle and apoptosis levels of DCs. Furthermore, enzyme‐linked immunosorbent assay was utilized to measure the levels of pro‐inflammatory cytokines (IL‐10, IL‐12 and IL‐23). The TLR4/NF‐κB signalling pathway in co‐cultured DCs was evaluated using the western blot assay. Through querying the Targetscan website, we predicted Rab27a as a target gene of miR‐124, which was subsequently validated through double luciferase assay, flow cytometry and other experimental approaches. Our findings demonstrate that MiR‐124 modulates the secretion of CRSwNP epithelial exosomes, thereby influencing DC function via its interaction with the target gene Rab27a. These results offer novel insights into potential therapeutic targets for the treatment of CRSwNP. [ABSTRACT FROM AUTHOR]

Objective: The most prevalent brain‐specific microRNA, MicroRNA‐124, exhibits anti‐inflammatory properties. Luteolin nano‐formulation with Zn oxide in the form of L/ZnO NPs may boost anti‐diabetic properties; however, its beneficial effect on miRNAs is yet unknown in diabetes. The effectiveness of L/ZnONPs supplements in preventing diabetic neurodegeneration by modulating inflammatory responses in a diabetic model was investigated. Methods: A diabetic rat model was induced by a high‐fat diet and streptozotocin (30 mg/kg I.P.). Plasma glucose, insulin, and HOMR‐IR levels, as well as cytokines, lipid peroxidation, GSH/GSSG, and glucose transporter 1, were determined along with the tight junction proteins occludin (OCLN) and zona occludens 1 (ZO‐1). Moreover, the expressions of brain CCAAT/enhancer‐binding protein (C/EBPA mRNA), miR‐124, glial fibrillary acidic protein (GFAP), and NF‐kBp65 were measured alongside the histological investigation. Results: The results revealed that L/ZnO NPs were able to diminish lipid peroxidation, increase the activity of antioxidant enzymes, and reduce inflammation under oxidative stress. Consequently, it was able to reduce hyperglycemia, elevate insulin levels, and improve insulin resistance. Besides, L/ZnO NPs upregulate miR‐124, reduce C/EBPA mRNA, increase BCl‐2, and inhibit apoptosis. The results indicate that diabetes raises BBB permeability via tight junction protein decline, which is restored following L/ZnO NPs treatment. Luteolin/ZnO NPs regulate miR‐124 and microglia polarization by targeting C/EBPA and are expected to alleviate inflammatory injury via modulation of the redox‐sensitive signal transduction pathways. Luteolin/ZnO NPs have a novel target for the protection of the BBB and the prevention of neurological complications in diabetes. [ABSTRACT FROM AUTHOR]

Cyprinid herpesvirus 2 (CyHV‐2), the etiological agent of herpesvirus haematopoietic necrosis (HVHN) in carp and goldfish, has caused significant economic losses in the aquaculture industry. During viral infection, the host initiates a series of active or passive defences to regulate the process of virus infection. Apoptosis is a key component of active cellular defence, and members of the Bcl‐2 family have been shown to play a critical role in the apoptotic process. However, the mechanism of action of the Bcl‐2 family in inducing apoptosis during CyHV‐2 infection remains unclear. In this study, we revealed the molecular mechanism of miRNA‐mediated silver crucian carp BAX (ccBax) in CyHV‐2‐induced apoptosis for the first time and demonstrated that the overexpression of miR‐124 suppressed ccBax expression and significantly down‐regulated apoptosis in caudal fin cells of Carassius auratus gibelio (GiCF), while miR‐124 inhibitors were the opposite. These studies indicated that miR‐124 inhibits CyHV‐2‐induced apoptosis by reducing the expression of ccBax. Furthermore, the fact that transfection of miR‐124 mimics promoted CyHV‐2 replication, whereas miR‐124 inhibitors inhibited CyHV‐2 replication, indicated that miR‐124 inhibited CyHV‐2‐induced apoptosis and contributed to viral replication. All these results suggested that miR‐124 suppresses virus‐induced apoptosis and promotes viral replication by targeting and regulating ccBax expression. [ABSTRACT FROM AUTHOR]

The activation of hepatic stellate cells is the primary function of facilitating liver fibrosis. Interfering with the coordinators of different signaling pathways in activated hepatic stellate cells (aHSCs) could be a potential approach in ameliorating liver fibrosis. Regarding the illustrated anti‐fibrotic effect of imatinib in liver fibrosis, we investigated the imatinib′s potential role in inhibiting HSC activation through miR‐124 and its interference with the STAT3/hepatic leukemia factor (HLF)/IL‐6 circuit. The anti‐fibrotic effect of imatinib was investigated in the LX‐2 cell line and carbon tetrachloride (CCl4)‐induced Sprague‐Dawley rat. The expression of IL‐6, STAT3, HLF, miR‐124, and α‐smooth muscle actin (α‐SMA) were quantified by quantitative real‐time PCR (qRT‐PCR) and the protein level of α‐SMA and STAT3 was measured by western blot analysis both in vitro and in vivo. The LX‐2 cells were subjected to immunocytochemistry (ICC) for α‐SMA expression. After administering imatinib in the liver fibrosis model, histopathological examinations were done, and hepatic function serum markers were checked. Imatinib administration alleviated mentioned liver fibrosis markers. The expression of miR‐124 was downregulated, while IL‐6/HLF/STAT3 circuit agents were upregulated in vitro and in vivo. Notably, imatinib intervention decreased the expression of IL‐6, STAT3, and HLF. Elevated expression of miR‐124 suppressed the expression of STAT3 and further inhibited HSCs activation. Our results demonstrated that imatinib not only ameliorated hepatic fibrosis through tyrosine kinase inhibitor (TKI) activity but also interfered with the miR‐124 and STAT3/HLF/IL‐6 pathway. Considering the important role of miR‐124 in regulating liver fibrosis and HSCs activation, imatinib may exert its anti‐fibrotic activity through miR‐124. [ABSTRACT FROM AUTHOR]

Ischaemic stroke (IS) is a cerebrovascular disease caused by cerebral infarction and cerebral artery occlusion. In this study, we proposed that EVs from bone marrow stromal cells (BMSCs) could reduce the impact of stroke by reducing the resultant glial cell activation and blood–brain barrier (BBB) leak. We furthermore investigated some of the signalling mechanisms. The transient middle cerebral artery occlusion (t‐MCAO) mouse model was established. The behavioural deficits and neuronal damage were verified using Bederson's scale and the 28‐point neurological score. The area of cerebral infarction was detected. The expressions of astrocytes/microglia markers and BBB permeability were evaluated by 2,3,5‐triphenyltetrazolium chloride (TTC) staining. The internalization of EVs by astrocytes/microglia in the peripheral area was detected by fluorescence labelling. The expressions of astrocyte/microglia markers were measured by RT‐qPCR. Levels of TNF‐α and IL‐1β in microglia were detected by ELISA. BBB permeability was evaluated. The downstream target genes and pathway of miR‐124 were analysed. Microglia/astrocytes were treated by oxygen–glucose deprivation reoxygenation (OGD/R). OGD/R microglia/astrocyte conditioned medium was used to culture bEnd.3 cells. The transendothelial electric resistance (TEER) of bEnd.3 cells was measured, and BBB permeability was characterized. Our results suggested that EVs from BMSCs can indeed reduce the extent of stroke‐mediated damage and evidenced that these effects are mediated via expression of the non‐coding RNA, miR‐124 that may act via the peroxiredoxin 1 (PRX1). Our results provided further motivation to pursue the use of modified EVs as a treatment option for neurological diseases. [ABSTRACT FROM AUTHOR]

Aims: In this study, the effect of intracerebral ventricle injection with a miR‐124‐3p agomir or antagomir on prognosis and on subventricular zone (SVZ) neural stem cells (NSCs) in adult rats with moderate traumatic brain injury (TBI) was investigated. Methods: Model rats with moderate controlled cortical impact (CCI) were established and verified as described previously. The dynamic changes in miR‐124‐3p and the status of NSCs in the SVZ were analyzed. To evaluate the effect of lateral ventricle injection with miR‐124‐3p analogs and inhibitors after TBI, modified neurological severity scores (mNSSs) and rotarod tests were used to assess motor function prognosis. The variation in SVZ NSC marker expression was also explored. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of predicted miR‐124‐3p targets was performed to infer miR‐124‐3p functions, and miR‐124‐3p effects on pivotal predicted targets were further explored. Results: Administration of miR‐124 inhibitors enhanced SVZ NSC proliferation and improved the motor function of TBI rats. Functional analysis of miR‐124 targets revealed high correlations between miR‐124 and neurotrophin signaling pathways, especially the TrkB downstream pathway. PI3K, Akt3, and Ras were found to be crucial miR‐124 targets and to be involved in most predicted functional pathways. Interference with miR‐124 expression in the lateral ventricle affected the PI3K/Akt3 and Ras pathways in the SVZ, and miR‐124 inhibitors intensified the potency of brain‐derived neurotrophic factor (BDNF) in SVZ NSC proliferation after TBI. Conclusion: Disrupting miR‐124 expression through lateral ventricle injection has beneficial effects on neuroregeneration and TBI prognosis. Moreover, the combined use of BDNF and miR‐124 inhibitors might lead to better outcomes in TBI than BDNF treatment alone. [ABSTRACT FROM AUTHOR]

Bcl2 family proteins play a critical role in cell death or survival. BAX, the death‐promoting protein of bcl2 family, mediated mitochondrial pathway inducing cells' apoptosis in mammal. MiRNAs have been implicated as negative regulators down‐regulating genes' expression after post‐transcriptional level. At present, little is known about the regulatory mechanism of miRNA on the Bcl2 family proteins during CyHV‐2 infection in silver crucian carp (Carassius auratus gibelio). In this study, the ccBAX (silver crucian carp BAX) gene was cloned and expressed, and polyclonal antibodies were raised in mouse against the purified ccBAX‐GST fusion protein. The structure analysis indicated that ccBAX protein included four conserve domains (BH1, BH2, BH3 and transmembrane domains) and the expression of ccBAX protein occurred throughout the cells. Furthermore, two miRNAs (miR‐124 and miRNA‐29b) were identified to negatively regulate ccBAX gene expression in GiCF cell. miR‐124 was found to suppress the expression of WT‐ccBAX (wild type), but not the MT‐ccBAX (mutant). Overall, the results demonstrated that the expression of the ccBAX gene was significantly down‐regulated by miR‐124 in silver crucian carp (Carassius auratus gibelio) during CyHV‐2 infection. [ABSTRACT FROM AUTHOR]

Incidence of intracerebral hemorrhage (ICH) and brain iron accumulation increases with age. Excess iron accumulation in brain tissues post‐ICH induces oxidative stress and neuronal damage. However, the mechanisms underlying iron deregulation in ICH, especially in the aged ICH model have not been well elucidated. Ferroportin1 (Fpn) is the only identified nonheme iron exporter in mammals to date. In our study, we reported that Fpn was significantly upregulated in perihematomal brain tissues of both aged ICH patients and mouse model. Fpn deficiency induced by injecting an adeno‐associated virus (AAV) overexpressing cre recombinase into aged Fpn‐floxed mice significantly worsened the symptoms post‐ICH, including hematoma volume, cell apoptosis, iron accumulation, and neurologic dysfunction. Meanwhile, aged mice pretreated with a virus overexpressing Fpn showed significant improvement of these symptoms. Additionally, based on prediction of website tools, expression level of potential miRNAs in ICH tissues and results of luciferase reporter assays, miR‐124 was identified to regulate Fpn expression post‐ICH. Higher serum miR‐124 levels were correlated with poor neurologic scores of aged ICH patients. Administration of miR‐124 antagomir enhanced Fpn expression and attenuated iron accumulation in aged mice model. Both apoptosis and ferroptosis, but not necroptosis, were regulated by miR‐124/Fpn signaling manipulation. Our study demonstrated the critical role of miR‐124/Fpn signaling in iron metabolism and neuronal death post‐ICH in aged murine model. Thus, Fpn upregulation or miR‐124 inhibition might be promising therapeutic approachs for this disease. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are known to regulate gene expression and modulate cellular differentiation. MicroRNA‐9/9* (miR‐9/9*) and microRNA‐124 (miR‐124) are highly expressed in the central nervous system. In vivo function of miR‐9/9* and miR‐124 has been investigated in detail, whereas there remain some discrepancies regarding neural development. To this end, we electroporated miR‐9/9*, miR‐124 or miR‐9/9*/124 expression plasmids into neonatal retinal progenitor cells (RPCs) in vivo and analyzed the fate of electroporated cells. Both miR‐9/9* and miR‐124 reduced the number of SOX9‐ and GS‐positive cells and increased that of TUBB3‐positive cells in the postnatal day 14 retina. No major effects on the proliferation and apoptosis of the electroporated cells were detected at least postnatal day 3. These indicated that miR‐9/9* and miR‐124 influence the cell fate of glial cells, thereby inducing their differentiation into neurons. Moreover, we found this cell fate modulation was occurred in RPCs indicating high‐level expression of miRNA, but not in the low level. Our results strongly suggest that high‐level miRNA overexpression is essential for directing cell fate by miR‐9/9* and miR‐124 interference. [ABSTRACT FROM AUTHOR]

Curcumin treatment was reported to delay the progression of OA, but its underlying mechanism remains unclear. In this study, we aimed to investigate the molecular mechanism underlying the role of curcumin in OA treatment. Accordingly, by conducting MTT and flow cytometry assays, we found that the exosomes derived from curcumin‐treated MSCs helped to maintain the viability while inhibiting the apoptosis of model OA cells. Additionally, quantitative real‐time PCR and Western blot assays showed that the exosomes derived from curcumin‐treated MSCs significantly restored the down‐regulated miR‐143 and miR‐124 expression as well as up‐regulated NF‐kB and ROCK1 expression in OA cells. Mechanistically, curcumin treatment decreased the DNA methylation of miR‐143 and miR‐124 promoters. In addition, the 3' UTRs of NF‐kB and ROCK1 were proven to contain the binding sites for miR‐143 and miR‐124, respectively. Therefore, the up‐regulation of miR‐143 and miR‐124 in cellular and mouse OA models treated with exosomes remarkably restored the normal expression of NF‐kB and ROCK1. Consequently, the progression of OA was attenuated by the exosomes. Our results clarified the molecular mechanism underlying the therapeutic role of MSC‐derived exosomes in OA treatment. [ABSTRACT FROM AUTHOR]

Chronic kidney disease (CKD) is a persistent kidney structural and functional disorder. Quercetin is one of active extracted flavonoids and has protective effects. Therefore, we proposed to survey the effect of Quercetin on CKD. HK‐2 cells were preprocessed by Quercetin and then irritated with lipopolysaccharide (LPS). CCK‐8 assay and flow cytometry were utilized to test viability and apoptosis. ELISA assay was utilized to estimate the IL‐6 and TNF‐α secretion. Western blot was performed to examine the expression of apoptosis and inflammation‐associated mediators. After that HK‐2 cells were transfected by miR‐124 inhibitor. And the above‐mentioned parameters were reassessed. LPS stimulated apoptosis and declined viability in HK‐2 cells. Additionally, LPS stimulated inflammatory responses. Meanwhile, Quercetin attenuated LPS‐stimulated apoptosis, production of IL‐6, and TNF‐α in experimental cells. Subsequently, MyD88 and miR‐124 expression was elevated by LPS and alleviated by Quercetin. Finally, Quercetin exerted its protective function through NF‐κB pathway via up‐regulating miR‐124. Our data demonstrated that Quercetin reduced apoptosis and inflammation stimulated by LPS in HK‐2 cells. Moreover, Quercetin alleviated LPS‐stimulated injury by up‐regulating miR‐124. [ABSTRACT FROM AUTHOR]

This study aimed to explore the roles of miR‐214 and MALAT1 rs619586 polymorphism in the control and survival of differentiated thyroid carcinoma (DTC) via Cox regression analyses. The levels of MALAT1, miR‐214, and CTNNB1 in different experimental groups were compared to study the interaction among MALAT1, miR‐214, and CTNNB1. MTT and colony assays were used to investigate the role of rs619586 polymorphism in cell growth. The G allele of rs619586 polymorphism obviously decreased the 5‐year survival of patients with DTC. Additionally, compared with AA‐genotyped patients, patients carrying the AG/GG genotypes of MALAT1 rs619586 polymorphism showed much higher levels of DTC grade and CTNNB1 expression, along with lower levels of MALAT1 and miR‐214 expression. Furthermore, the transcription activity of MALAT1 was significantly lowered by the rs619586G allele or miR‐214 mimic, while the miR‐214 inhibitor upregulated the luciferase activity of MALAT1. Additionally, miR‐214 inhibited CTNNB1 expression by targeting CTNNB1 3'‐untranslated region. Finally, the G allele of MALAT1 rs619586 polymorphism apparently promoted cell proliferation. Our study indicated that miR‐214 inhibited MALAT1 expression by directly binding to the G allele of MALAT1 rs619586 polymorphism, thus inhibiting CTNNB1 expression and promoting cell proliferation in the pathogenesis of DTC. Therefore, MALAT1 rs619586 polymorphism could be used to predict the prognosis of DTC. [ABSTRACT FROM AUTHOR]

Alzheimer's disease (AD) is a chronic progressive neurodegenerative disorder. However, its pathogenetic mechanism is still poorly understood. An increasing number of studies have evidenced the important role of long noncoding RNAs (lncRNAs) in AD. The aim of the current study was to investigate the effect and molecular mechanism of the lncRNA X‐inactive specific transcript (XIST) in AD. Bilateral common carotid artery occlusion (2VO) was used to induce an AD model in mice. Hydrogen peroxide (H2O2) was used to induce an AD model in N2a cells. The lncRNA XIST, miR‐124, and BACE1 messenger RNA expression levels were detected by a real‐time polymerase chain reaction. The BACE1 protein expression level was detected by western blot and immunofluorescence assay. The Aβ1–42 expression level was detected using an enzyme‐linked immunosorbent assay kit. The expression level of lncRNA XIST was significantly upregulated in AD models, both in vivo and in vitro. Silencing of lncRNA XIST negatively regulated miR‐124 and positively regulated BACE1 expression in N2a cells, which is attenuated by cotransfection of anti‐miR‐124 oligodeoxyribonucleotide (AMO‐124). Silencing of lncRNA XIST reversed the effect of H2O2 on miR‐124, BACE1, and Aβ1–42 expression in N2a cells, which was reversed by cotransfection of AMO‐124. Silencing of lncRNA XIST attenuated AD‐related BACE1 alteration through miR‐124. LncRNA XIST may be a new potential target for the treatment of AD. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are noncoding single‐stranded RNAs, approximately 20‐24 nucleotides in length, known as powerful posttranscriptional regulators. miRNAs play important regulatory roles in cellular processes by changing messenger RNA expression and are widely involved in human diseases, including tumors. It has been reported in the literature that miRNAs have a precise role in cell proliferation, programmed cell death, differentiation, and expression of coding genes. MicroRNA‐124 (miR‐124) has reduced exparession in various human neoplasms and is believed to be related to the occurrence, development, and prognosis of malignant tumors. In our review, we focus on the specific molecular functions of miR‐124 and the downstream gene targets in major cancers, which provide preclinical evidence for the treatment of human cancer. Although some obstacles exist, miR‐124 is still attracting intensive research focus as a promising and effective anticancer weapon. [ABSTRACT FROM AUTHOR]

Backgrounds: Necrotizing enterocolitis (NEC) was one of the main causes of morbidity and mortality in neonates. Our objective was to detect the mechanism of miR‐124 in small bowel tissues of NEC. Methods: Hematoxylin and eosin (H&E) staining was used to detect the repair of the damaged tissues in rat NEC model. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was used to evaluate the cell apoptosis level in intestinal tissue. Reverse transcription polymerase chain reaction (RT‐PCR) was used to detect the messenger RNA (mRNA) expression level of miR‐124, Rho‐associated coiled‐coil‐containing protein kinase 1 (ROCK1), myosin phosphatase target subunit 1 (MYPT1), and Toll‐like receptor 9 (TLR9) in NEC tissues and IEC‐6 cells. Luciferase reporter assay was used to verify whether ROCK1 is a direct target of miR‐124. Results: miR‐124 was overexpressed in the NEC tissues, while ROCK1 and MYPT1 was downregulated in the NEC tissues. Inhibition of miR‐124, suppressed the intestinal cell apoptosis and promoted the expression of ROCK1 and MYPT1. What is more, overexpression of miR‐124 could inhibit the expression of ROCK1, TLR9, and MYPT1. Luciferase assay confirmed that miR‐124 can regulate the transcriptional activity of ROCK1 through binding its 3′‐UTR region. Conclusion: miR‐124 was a promoter of NEC, which promotes the intestine cell apoptosis and inflammatory cell infiltration through the inhibition of TLR9 expression by targeting ROCK1. miR‐124 was a promoter of necrotizing enterocolitis (NEC), which promotes the intestine cell apoptosis and inflammatory cell infiltration through the inhibition of Toll‐like receptor 9 (TLR9) expression by targeting rho‐associated coiled‐coil‐containing protein kinase 1 (ROCK1). [ABSTRACT FROM AUTHOR]

Hair follicle stem cells (HFSCs) are able to differentiate into neurons and glial cells. Distinct microRNAs (miRNAs) regulate the proliferation and differentiation of HFSCs. However, the exact role of miR‐124 in the neural differentiation of HFSCs has not been elucidated. HFSCs were isolated from mouse whisker follicles. miR‐9, let‐7b, and miR‐124, Ptbp1 , and Sox9 expression levels were detected by real‐time polymerase chain reaction (RT‐PCR). The influence of miR‐124 transfection was evaluated using immunostaining. We demonstrated that miR‐124 and let‐7b expression levels were significantly increased after the neural differentiation. Sox9 and Ptbp1 were identified as the target of miR‐124 in the HFSCs. During neural differentiation and miR‐124 mimicking, Ptbp1 and Sox9 levels were decreased. Moreover, the miR‐124 overexpression increased MAP2 (58.43 ± 11.26) and NeuN (48.34 ± 11.15) proteins expression. The results demonstrated that miR‐124 may promote the differentiation of HFSCs into neuronal cells by targeting Sox9 and Ptbp1. miR‐124 and let‐7b are upregulated in hair follicle bulge cells after neural differentiation. Upregulation of miR‐124 triggers neural differentiation. Ptbp1 and Sox9 mRNAs are direct miR‐124 targets during neural differentiation of hair follicle stem cells (HFSCs). [ABSTRACT FROM AUTHOR]

Metastasis is the primary contributor to colorectal cancer mortality. High‐intensity focused ultrasound (HIFU) is an emerging technology for tumor therapy that exerts its effects through tumor ablation, mechanical disruption, and enhancement of immune responses. However, it remains unclear whether HIFU can influence tumor metastasis. Here, we examined the effect of HIFU on tumor metastasis of colorectal cancer cells and the underlying mechanisms. HIFU was observed to inhibit migration of HCT‐116 cells in vitro and suppress lung metastasis in a mouse model of colon cancer. In addition, HIFU up‐regulated microRNA (miR) ‐124 expression, which inhibited the activation of signal transducer and activator of transcription 3 (STAT3) and inhibited migration of HCT‐116 cells. Treatment with an inhibitor of miR‐124 reversed the effect of HIFU on cell migration. In conclusion, our results suggest that HIFU exerts anti‐metastatic effects in colon cancer, and this effect is possibly mediated via up‐regulation of miR‐124 and subsequent miR‐124‐mediated STAT3 suppression. [ABSTRACT FROM AUTHOR]

Bone repair involves bone resorption through osteoclastogenesis and the stimulation of neovascularization and osteogenesis by endothelial progenitor cells (EPCs). However, the role of EPCs in osteoclastogenesis is unclear. In this study, we assess the effects of EPC‐derived exosomes on the migration and osteoclastic differentiation of primary mouse bone marrow‐derived macrophages (BMMs) in vitro using immunofluorescence, western blotting, RT‐PCR and Transwell assays. We also evaluated the effects of EPC‐derived exosomes on the homing and osteoclastic differentiation of transplanted BMMs in a mouse bone fracture model in vivo. We found that EPCs cultured with BMMs secreted exosomes into the medium and, compared with EPCs, exosomes had a higher expression level of LncRNA‐MALAT1. We confirmed that LncRNA‐MALAT1 directly binds to miR‐124 to negatively control miR‐124 activity. Moreover, overexpression of miR‐124 could reverse the migration and osteoclastic differentiation of BMMs induced by EPC‐derived exosomes. A dual‐luciferase reporter assay indicated that the integrin ITGB1 is the target of miR‐124. Mice treated with EPC‐derived exosome‐BMM co‐transplantations exhibited increased neovascularization at the fracture site and enhanced fracture healing compared with those treated with BMMs alone. Overall, our results suggest that EPC‐derived exosomes can promote bone repair by enhancing recruitment and differentiation of osteoclast precursors through LncRNA‐MALAT1. [ABSTRACT FROM AUTHOR]

Aim: To understand the mechanism of long non‐coding RNA (LncRNA) HOTAIR on renal interstitial fibrosis (RIF) by regulating Notch1 pathway via the modulation of miR‐124. Methods: Unilateral ureteral occlusion (UUO) was used to construct the RIF rat model. HK‐2 cells induced by TGF‐β1 were used for the in vitro experiment, which were divided into five groups: Vehicle, TGF‐β1, si‐HOTAIR+TGF‐β1, miR‐124 inhibitor+TGF‐β1, and si‐HOTAIR+miR‐124 inhibitor+TGF‐β1 groups. Quantitative real‐time PCR (qRT‐PCR) and Western blot were performed to detect the expression of HOTAIR, miR‐124, Notch1‐ and epithelial‐to‐mesenchymal transition (EMT)‐related proteins. Results: Significant elevated HOTAIR and reduced miR‐124 were presented in UUO rats and TGF‐β1‐induced HK‐2 cells in a time‐dependent manner, with the increased Jagged1 (JAG1), Notch1, NICD, α‐SMA and FN, as well as the decreased E‐cadherin (all P < 0.05). Compared with the TGF‐β1 group, cells in the si‐HOTAIR+TGF‐β1 group were remarkably declined in cell proliferation and the protein expressions of JAG1, Notch1, NICD, α‐SMA, and FN, but dramatically higher in E‐cadherin expression (all P < 0.05). However, in comparison with the si‐HOTAIR+TGF‐β1 group, cells in the si‐HOTAIR+miR‐124 inhibitor+TGF‐β1 group were apparently improved in proliferation and the protein expression of JAG1, Notch1, NICD, α‐SMA, and FN, but substantially reduced in the level of E‐cadherin protein (all P < 0.05). Conclusion: Silencing lncRNA HOTAIR can up‐regulate miR‐124 to block Notch1 pathway, and thereby alleviating EMT and RIF, indicating HOTAIR as a potential target for RIF treatment. Summary at a Glance: This study demonstrates elevated levels of HOTAIR in mouse kidneys with interstitial fibrosis. It also shows that silencing lncRNA HOTAIR upregulates miR‐124 to block the Notch1 pathway. This alleviates TGF‐β induced fibrotic mechanisms in cultured human proximal tubular cells, suggesting HOTAIR is a potential target for treating kidney interstitial fibrosis. [ABSTRACT FROM AUTHOR]

lncRNAs can exert many biological effects in several cancer types. MALAT1 is a kind of lncRNA which is greatly overexpressed in several tumors including non-small cell lung cancer (NSCLC). However, the mechanism of MALAT1 in NSCLC still remains unclear. In our current study, we concentrated on the biological mechanism of MALAT1 in NSCLC. It was observed that MALAT1 was significantly upregulated in five human NSCLC cells including A549, H23, H522, H1299, and H460 cells compared to normal bronchial epithelial cell line 16HBE cells. On the contrary, miR-124 was remarkably downregulated, which indicated a potential negative correlation between miR-124 and MALAT1. MALAT1 inhibition can increase miR-124 expression in A549 and H460 cells. In addition, miR-124 mimics were able to repress MALAT1 expression and miR124 inhibitors can promote MALAT1 levels. Then it was found that shMALAT1 can inhibit NSCLC cell proliferation, colony formation and apoptosis, which can be reversed by miR-124 inhibitors. Bioinformatic analysis predicted the correlation between miR-124 and MALAT1. In addition, STAT3 was found to be a novel mRNA target of miR-124. Downregulation of MALAT1 can inhibit NSCLC development by enhancing miR-124 and decreasing STAT3 expression. We speculated that MALAT1can act as a competing endogenous lncRNA (ceRNA) to modulate miR-124/STAT3 in NSCLC. Taken these together, we revealed that MALAT1/miR-124/STAT3 was involved in NSCLC development. [ABSTRACT FROM AUTHOR]

The class IIa histone deacetylases (HDACs) play important roles in the central nervous system during diverse biological processes such as synaptic plasticity, axon regeneration, cell apoptosis, and neural differentiation. Although it is known that HDAC5 regulates neuronal differentiation, neither the physiological function nor the regulation of HDAC5 in neuronal differentiation is clear. Here, we identify HDAC5 as an inhibitor of neurite elongation and show that HDAC5 is regulated by the brain enriched microRNA miR-124 and miR-9. We discover that HDAC5 inhibits neurite extension both in differentiated P19 cells and primary neurons. We also show that the neuronal membrane glycoprotein GPM6A (M6a) is a direct target gene of HDAC5 regulated transcriptional factor MEF2C. HDAC5 inhibits neurite elongation, acting at least partially via a MEF2C/M6a signaling pathway. We also confirmed the miR-124/miR-9 regulated HDAC5-MEF2C-M6a pathway regulates neurite development in primary neurons. Thus, HDAC5 emerges as a cellular conductor of MEF2C and M6a activity and is regulated by miR-124 and miR-9 to control neurite development. [ABSTRACT FROM AUTHOR]

S100 calcium-binding protein A4 (S100A4) induces proliferation and migration of vascular smooth muscle cells ( VSMCs). We aimed to find the micro RNA regulating S100A4 expression. S100A4 transcripts are abruptly increased in the acute phase of carotid arterial injury 1 day later (at day 1) but gradually decreases at days 7 and 14. Bioinformatics analysis reveals that miR-124 targets S100A4. VSMC survival is attenuated by miR-124 mimic but increased by miR-124 inhibitor. miR-124 decreases immediately after carotid arterial injury but dramatically increases at days 7 and 14. miR-124 inhibitor-induced cell proliferation is blocked by S100A4 si RNA, whereas miR-124-induced cell death is recovered by S100A4. Our findings suggest that miR-124 is a novel regulator of VSMC proliferation and may play a role in the development of neointimal proliferation. [ABSTRACT FROM AUTHOR]

Long noncoding RNAs (lnc RNAs) have recently emerged as pivotal regulators in governing fundamental biological processes, as well as in tumorigenesis. The nuclear paraspeckle assembly transcript 1 ( NEAT1) is one of the most highly regulated lnc RNAs in recent genomic datasets, however, its biological role and regulatory mechanism in ovarian cancer ( OC) development and progression are poorly defined. In this study, we identified that NEAT1 was up-regulated in OC patients and cell lines, and its expression was associated with the FIGO stage and lymph node metastasis. Furthermore, the ectopic expression of NEAT1_1 in OVCAR-3 cell lines promoted cell proliferation and invasion, whereas knockdown of NEAT1_1 did the opposite. Furthermore, NEAT1_1 was stabilized by an RNA-binding protein HuR, but suppressed by miR-124-3p in OC cells. Accordingly, the increased HuR mRNA and decreased miR-124-3p levels were observed in OC patients. These results suggested that lnc RNA NEAT1, whose expression was collaboratively controlled by HuR and miR-124-3p, could regulate ovarian carcinogenesis and may serve as a potential target for antineoplastic therapies. [ABSTRACT FROM AUTHOR]

Cancer cells in hypoxia usually make adaptive changes in cellular metabolism, such as altered autophagy. This might be a cause of enhanced radioresistance in some types of cancer. In this study, we investigated hypoxia-responsive mi RNAs in two prostate cancer cell lines ( DU145 and PC3). This study firstly reported that hypoxia induces further downregulation of miR-124 and miR-144, which might be a result of impaired dicer expression. These two mi RNAs can simultaneously target 3′ UTR of PIM1. Functional study showed that miR-124 or miR-144 overexpression can inhibit hypoxia-induced autophagy and enhance radiosensitivity at least via downregulating PIM1. Therefore, hypoxia induced miR-124 and miR-144 downregulation may contribute to a prosurvival mechanism of prostate cancer cells to hypoxia and irradiation at least through attenuated suppressing of PIM1. This finding presents a potential therapeutic target for prostate cancer. [ABSTRACT FROM AUTHOR]

Parkinson's disease ( PD) is the most prevalent movement disorder characterized by selective loss of midbrain dopaminergic ( DA) neurons. MicroRNA-124 (mi R-124) is abundantly expressed in the DA neurons and its expression level decreases in the 1-methyl-4-pheny-1, 2, 3, 6-tetrahydropyridine ( MPTP) model of PD. However, whether the upregulation of miR-124 could attenuate neurodegeneration remains unknown. Here, we employed miR-124 agomir and miR-124 mimics to upregulate mi R-124 expression in MPTP-treated mice and MPP+-intoxicated SH- SY5Y cells, respectively. We found that loss of DA neurons and striatal dopamine in MPTP-treated mice was significantly reduced by upregulating mi R-124. In addition, we identified a target of mi R-124, Bim that mediated the neuroprotection of mi R-124. Indeed, treatment of mi R-124 agomir in MPTP-treated mice inhibited Bim expression, thus suppressing Bax translocation to mitochondria. Moreover, impaired autophagy process in MPTP-treated mice and MPP+-intoxicated SH- SY5Y cells characterized as autophagosomes ( AP) accumulation and lysosomal depletion were alleviated by the upregulation of miR-124. Taken together, these results indicate that upregulation of mi R-124 could regulate apoptosis and impaired autophagy process in the MPTP model of PD, thus reducing the loss of DA neurons. [ABSTRACT FROM AUTHOR]

Aberrant Notch signalling plays an important role in cancer progression. However, little is known about the interaction between mi RNA and the Notch signalling pathway and its role in gastric cancer ( GC). In this study, we found that miR-124 was down-regulated in GC compared with adjacent normal tissue. Forced expression of miR-124 inhibited GC cell growth, migration and invasion, and induced cell cycle arrest. miR-124 negatively regulated Notch1 signalling by targeting JAG1. miR-124 levels were also shown to be inversely correlated with JAG1 expression in GC. Furthermore, we found that the overexpression of the intracellular domain of Notch1 repressed miR-124 expression, promoted GC cell growth, migration and invasion. Conversely, blocking Notch1 using a γ-secretase inhibitor up-regulated miR-124 expression, inhibited GC cell growth, migration and invasion. In conclusion, our data demonstrates a regulatory feedback loop between miR-124 and Notch1 signalling in GC cells, suggesting that the miR-124/Notch axis may be a potential therapeutic target against GC. [ABSTRACT FROM AUTHOR]

UHRF1, an epigenetic factor, is implicated in various cellular processes of tumorigenesis. However, the modulation of UHRF1 expression in human bladder cancer at post-transcriptional levels remains unclear. Here, we report that miR-124 suppresses expression of UHRF1 to affect the progression of human bladder cancer through competitive binding of the same region of its 3'-UTR. We show that compared with corresponding normal tissues, UHRF1 is upregulated and miR-124 is downregulated in bladder cancer tissues, demonstrating an inverse correlation of miR-124 and UHRF1. Quantitative PCR and western blot assay demonstrated that over-expression of miR-124 resulted in the suppression of UHRF1. Furthermore, luciferase assay revealed that miR-124 could control the fate of target gene UHRF1 mRNA by binding 3'-UTR. The rescue experiment confirmed that miR-124 exerted its biological functions by targeting UHRF1. miR-124 over-expression significantly attenuated cellular proliferation, migration, invasion and vasculogenic mimicry in vitro, and tumor growth in vivo. UHRF1 siRNA showed significant inhibitory effects on bladder cancer cells. Collectively, our study demonstrates that miR-124 can impair the proliferation or metastasis of human bladder cancer cells by down-regulation of UHRF1. [ABSTRACT FROM AUTHOR]

Neurodegenerative diseases are one of the most challenging subjects in medicine. Investigation of their underlying genetic or epigenetic factors is hampered by lack of suitable models. Patient-specific induced pluripotent stem cells ( iPS cells) represent a valuable approach to provide a proper model for poorly understood mechanisms of neuronal diseases and the related drug screenings. miR-124 and miR-128 are the two brain-enriched mi RNAs with different time-points of expression during neuronal development. Herein, we transduced human iPS cells with miR-124 and miR-128 harboring lentiviruses sequentially. The transduced plasmids contained GFP and puromycin antibiotic-resistant genes for easier selection and identification. Morphological assessment and immunocytochemistry (overexpressions of beta-tubulin and neuron-specific enolase) confirmed that induced hi PS cells by miR-124 and miR-128 represent similar characteristics to those of mature neurons. In addition, the upregulation of neuron-specific enolase, beta-tubulin, Map2, GFAP, and BDNF was detected by quantitative real-time PCR. In conclusion, it seems that our novel protocol remarks the combinatorial effect of miR-124 and miR-128 on neural differentiation in the absence of any extrinsic factor. Moreover, such cellular models could be used in personalized drug screening and applied for more effective therapies. [ABSTRACT FROM AUTHOR]

MicroRNA-124 (miR-124) is an evolutionarily conserved, small, noncoding RNA molecule that participates in the central nervous system (CNS) developmental control of gene expression. In the current study, we found that Drosophila embryos lacking the mir-124 gene did not exhibit detectable defects in axon growth or CNS development. On the other hand, adult mutants showed severe problems in locomotion, flight, and female fertility. Furthermore, the deficits that we observed in the adult stage could be marginally rescued with elav-GAL4 driven expression of miR-124, making elav-GAL4 an excellently simulated driver to induce entopic over-expression of miR-124. Further developmental assessment in the third instar larval neuromuscular junction (NMJ) and dendritic arborization (DA) neurons was performed with miR-124 knock outs, flies over-expressing miR-124, and rescue models. Typically, the absence and over-abundance of a molecular signal exerts opposite effects on development or phenotype. However, we determined that both miR-124 knock-outs and over-expressing flies displayed reduced NMJ 6/7 bouton number and branch length. Similarly, reduced ddaE branching numbers were observed between the two mutant lines. As to ddaF, we found that branching number was not influenced by mir-124 knock out, but was statistically reduced by miR-124 over-expression. While we were not able to determine any causal relationship between behavioral defects and dysplasia of NMJs or DA neurons, there were some accompanying relationships among behavioral phenotypes, NMJ abnormalities, and ddaE/ddaF dendritic branching which were all controlled by miR-124. [ABSTRACT FROM AUTHOR]

Highlights: [•] CLOCK expression is upregulated in high-grade glioma tissues vs. low-grade or normal tissues. [•] MiR-124 can directly target and repress Clock. [•] The upregulation of CLOCK is likely a consequence of the downregulation of miR-124 in gliomas. [•] CLOCK can positively regulate glioma proliferation and migration. [•] The effects of CLOCK on tumorigenesis are likely the result of enhanced NF-κB signaling. [Copyright &y& Elsevier]

Gene expression in the rodent brain is dependent on exercise. However, our findings regarding this fact are inadequate. The aim of this study was to evaluate the effect of exercise intensity on changes in factors involved in neuronal differentiation and plasticity in the hippocampus of adult male rats. Thus, the effects of 2 weeks treadmill running at of low (11 m/min) and high (25 m/min) intensities, 30 min/day for 14 consecutive days on the expression of two genes, microRNA-124 and neuron-restrictive silencer factor/RE1-silencing transcription factor (REST) as well as changes of two proteins of brain-derived neurotrophic factor (BDNF) and tyrosine kinase B (TrkB) were analysed. Using quantitative real-time PCR techniques have shown that both exercise intensities reduced the expression of REST (31% in low exercise intensity, 52% in high exercise intensity) and elevated the expression of miR-124 (3.8-fold in low exercise intensity, 11.9-fold in high exercise intensity) significantly ( P ≤ 0.05). This suggests that exercise probably through the changes in these genes is involved in neuronal differentiation in the hippocampus of rats. BDNF and TrkB levels were measured by ELISA. Exercise, at low intensity, increased TrkB (4.51 ng/L vs. 3.73 ng/L) and BDNF (11.97 ng/L vs. 11.08 ng/L) proteins significantly ( P ≤ 0.05), while at high intensity, the changes in comparison with the control group were not significant. Expression levels during the exercise programme with high intensity were dramatic for miR-124 and less dramatic for REST compared with low intensity exercise programme. [ABSTRACT FROM AUTHOR]

Abstract: Hepatitis C Virus core protein (HCVc) plays important roles in the development of intrahepatic cholangiocarcinoma (ICC). MicroRNAs (miRNAs) contribute to tumor progression by interacting with downstream target genes. However, the regulation and role of miRNAs in HCV-related intrahepatic cholangiocarcinoma (HCV-ICC) is poorly understood. In this study, we found that miR-124 was down-regulated in HCV-ICC and the induction of DNMT1 by HCVc mediated the suppression of miR-124. Over-expression of miR-124 suppressed cell migration and invasion in vitro, and reduced the protein levels of SMYD3 and downstream target genes (c-Myc and MMP9). Knockdown of SMYD3 inhibited cell migration and invasion resembling that of miR-124 over-expression. In conclusion, our studies indicate that low miR-124 levels mediated by HCVc via DNMT1 promote ICC cell migration and invasion by targeting SMYD3. [Copyright &y& Elsevier]

SPHK1 expression is elevated in gastric cancer and is associated with shorter survival times for patients. However, the molecular mechanism of SPHK1 up-regulation in gastric cancer remains unclear. In the present study, we report that miR-124 down-regulated SPHK1 expression by directly targeting its 3′-untranslated region (3′-UTR) and that miR-124 expression was inversely correlated with SPHK1 expression in gastric cancer samples. Furthermore, we demonstrated that, similar to the effect of silencing SPHK1, up-regulation of miR-124 markedly inhibited proliferation and tumourigenicity of gastric cancer cells both in vitro and in vivo. This was found to be mechanistically associated with induction of cyclin-dependent kinase inhibitors p21 $^{{\rm Cip1}}$ and p27 $^{{\rm Kip1}}$, enhancement of the transcriptional activity of FOXO1 and suppression of AKT activity. Moreover, we showed that the re-introduction of SPHK1 (without the 3′-UTR), but not with the 3′-UTR, could abrogate the miR-124-mediated induction of p21 $^{{\rm Cip1}}$ and p27 $^{{\rm Kip1}}$, as well as rescue the miR-124-induced proliferation inhibition. Together, these results suggest that miR-124 has an important role in the suppression of gastric cancer and presents a novel mechanism of miRNA-mediated SPHK1 expression in cancer cells. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Abstract: Neuroblastoma is the most common extracranial solid tumor in children. We investigate whether miR-124, the abundant neuronal miRNA, plays a pivotal role in neuroblastoma. Knockdown of miR-124 promotes neuroblastoma SK-N-SH cell differentiation, cell cycle arrest and apoptosis. Further miR-124 is predicted to target aryl hydrocarbon receptor (AHR) which may promote neuroblastoma cell differentiation. We validate that miR-124 may suppress the expression of AHR by targeting its 3′-UTR. These results suggest that miR-124 could serve as a potential therapeutic target of neuroblastoma. [Copyright &y& Elsevier]

The β-amyloid peptide that accumulate in Alzheimer's disease (AD) brain derive from proteolytic processing of the amyloid precursor protein (APP). Recent evidence suggest that microRNAs (miRNAs) participate in the post-transcriptional regulation of APP expression. Because gene dosage effects of the APP gene can cause genetic AD, dysregulation of the miRNA network could contribute significantly to disease. Here, we present evidence that, besides APP expression regulation, miRNAs are equally involved in the regulation of neuronal APP mRNA alternative splicing. Lack of miRNAs in postmitotic neurons in vivo is associated with APP exons 7 and 8 inclusion, while ectopic expression of miR-124, an abundant neuronal-specific miRNA, reversed these effects in cultured neurons. Similar results were obtained by depletion of endogenous polypyrimidine tract binding protein 1 (PTBP1) in cells, a recognized miR-124 target gene. Furthermore, PTBP1 levels correlate with the presence of APP exons 7 and 8, while PTBP2 levels correlate with the skipping of these exons during neuronal differentiation. Finally, we show that miR-124 is down-regulated in AD brain. In sum, our results suggest that specific miRNAs are involved in the fine-tuning of APP alternative splicing in neurons. Since abnormal neuronal splicing of APP affects β-amyloid peptide production, these results could contribute to the understanding of the implication of miRNAs in brain health and disease. [ABSTRACT FROM AUTHOR]

Abstract: Alterations in the levels of molecules which interact with the extracellular matrix, such as integrins, are associated with invasion of oral squamous cell carcinomas (OSCC). The molecular mechanisms underlying dysregulation of integrin expression in OSCC, however, remain unclear. Here, we show that microRNA-124, a small non-coding RNA down-regulated in OSCC, is able to downregulate expression of integrin beta-1 (ITGB1) by interacting with its 3′ untranslated region. Over-expression of miR-124 attenuates endogenous ITGB1 expression and reduces the adherence and motility of OSCC cells, suggesting disruption of miR-124-mediated repression of ITGB1 may be a key factor in OSCC progression. [ABSTRACT FROM AUTHOR]

Whether epigenetic factors such as DNA methylation and microRNAs interact to control adult hippocampal neurogenesis is not fully understood. Here, we show that Down syndrome critical region 1 (DSCR1) protein plays a key role in adult hippocampal neurogenesis by modulating two epigenetic factors: TET1 and miR‐124. We find that DSCR1 mutant mice have impaired adult hippocampal neurogenesis. DSCR1 binds to TET1 introns to regulate splicing of TET1, thereby modulating TET1 level. Furthermore, TET1 controls the demethylation of the miRNA‐124 promoter to modulate miR‐124 expression. Correcting the level of TET1 in DSCR1 knockout mice is sufficient to prevent defective adult neurogenesis. Importantly, restoring DSCR1 level in a Down syndrome mouse model effectively rescued adult neurogenesis and learning and memory deficits. Our study reveals that DSCR1 plays a critical upstream role in epigenetic regulation of adult neurogenesis and provides insights into potential therapeutic strategy for treating cognitive defects in Down syndrome. Synopsis: The Down syndrome critical region 1 (DSCR1) protein, which is upregulated in Down syndrome, plays a significant role in adult hippocampal neurogenesis. This work provides evidence that DSCR1 acts upstream of two epigenetic factors, TET1 and miRNA‐124, and that interplay between DNA methylation and miRNA pathways regulate adult neurogenesis and learning and memory. DSCR1 knockout and transgenic brains show defective hippocampal adult neurogenesis.DSCR1 binds to the introns of TET1, which regulates TET1mRNA splicing and its expression.TET1 regulates DNA methylation in the promoter of miRNA‐124, which subsequently alters miR‐124 expression.Defects both in adult neurogenesis and learning & memory manifested in Ts65Dn are prevented by reducing triplicate DSCR1 to duplicate in Ts65Dn. [ABSTRACT FROM AUTHOR]