RESUMEN
miR-124 is ubiquitously expressed in the nervous tissue and acts as a negative regulator of neuroinflammation. In the present study, we analyzed the possible role of miR-124 in response to LPS in the human microglial cell line. Our data revealed that the miR-124 anti-inflammatory effect is based not only on the suppression of MyD88 - NFκB pathway and downregulation of pro-inflammatory cytokines IL-1ß and IL-6 but also on the enhancement of TRAM-TRIF signaling and increased IFN-ß expression. Furthermore, the NFκB reporter assay demonstrated that specific miR-124 - induced NFκB activity changes could be detected only using NFκB reporter promoters lacking ATF/CREB binding site.
Asunto(s)
Interferón beta/genética , MicroARNs/metabolismo , Microglía/inmunología , Línea Celular , Humanos , Interferón beta/metabolismo , Receptores de Lipopolisacáridos/genética , Receptores de Lipopolisacáridos/metabolismo , Lipopolisacáridos/inmunología , MicroARNs/agonistas , Microglía/metabolismo , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Transducción de Señal/inmunología , Receptor Toll-Like 4/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/inmunologíaRESUMEN
Tuberculosis (TB) is caused by Mycobacterium tuberculosis (M. tuberculosis) infection and has the highest mortality rate of any single infectious disease worldwide. The aim of the present study was to investigate the function of microRNA (miR)5023p in M. tuberculosisinfected macrophages. The Gene Expression Omnibus database was used to analyze miR5023p expression in patients with TB and healthy individuals. THP1 and RAW 264.7 cells were transfected with miR5023p mimic, miR5023p inhibitor, pcDNA3.1ROCK1 or their negative controls. The expression levels of miR5023p and inflammatory cytokines were evaluated using reverse transcriptionquantitative PCR. The colonyforming unit assay was performed to assess the survival of M. tuberculosis in macrophages, and Tolllike receptor (TLR)4/NFκB signaling pathwayassociated protein expression levels were detected by western blotting. The nuclear translocation of NFκB p65 was detected via immunocytochemistry. TargetScan was used to predict the binding sites between miR5023p and ROCK1. The interaction between miR5023p and Rhoassociated coiledcoilforming protein kinase 1 (ROCK1) was confirmed using a dualluciferase reporter assay; ROCK1 was demonstrated to be a direct target gene of miR5023p. Results from the present study demonstrated that miR5023p expression was significantly increased during M. tuberculosis infection in macrophages. Upregulation of miR5023p expression levels significantly enhanced the survival of intracellular M. tuberculosis. IL6, TNFα, and IL1ß mRNA expression levels were significantly upregulated during M. tuberculosis infection but were downregulated by miR5023p overexpression. Moreover, miR5023p mimics transfection significantly downregulated TLR4/NFκB signaling pathwayassociated protein expression and significantly reduced nuclear transcription of NFκB in M. tuberculosisinfected macrophages. ROCK1 overexpression reversed the miR5023p inhibitory effect on cytokine production in M. tuberculosisinfected macrophages. In conclusion, miR5023p/ROCK1 may serve an antiinflammatory role and may improve the survival of M. tuberculosis within macrophages, which may provide a promising therapeutic target for TB.
Asunto(s)
Macrófagos/inmunología , MicroARNs/metabolismo , Mycobacterium tuberculosis/inmunología , Tuberculosis/genética , Quinasas Asociadas a rho/genética , Adolescente , Adulto , Animales , Estudios de Casos y Controles , Línea Celular Tumoral , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/inmunología , Femenino , Voluntarios Sanos , Interacciones Huésped-Patógeno/efectos de los fármacos , Interacciones Huésped-Patógeno/genética , Humanos , Macrófagos/metabolismo , Macrófagos/microbiología , Masculino , Ratones , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , Persona de Mediana Edad , Mycobacterium tuberculosis/aislamiento & purificación , Células RAW 264.7 , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Transducción de Señal/inmunología , Receptor Toll-Like 4/metabolismo , Factor de Transcripción ReIA/metabolismo , Tuberculosis/sangre , Tuberculosis/inmunología , Tuberculosis/microbiología , Adulto Joven , Quinasas Asociadas a rho/metabolismoRESUMEN
Tumor ulceration is considered one of the most prognostically significant findings in primary cutaneous melanoma, associated with decreased disease-free and overall survival. However, the unique features associated with ulcerated melanoma that contribute to a poor prognosis in affected patients remain poorly defined. microRNAs are small, non-coding RNAs that function to inhibit expression of specific gene targets, therefore altering the functions of cells in which they are expressed. miR-1469 is a novel miR with significantly decreased expression in ulcerated melanoma tissue relative to non-ulcerated tumors. We hypothesized that loss of miR-1469 expression in melanoma contributes to altered tumor cell functions mediating disease progression. Transfection of a miR-1469 mimic resulted in a significant reduction in the migratory and invasive capacity of the CHL1 and MEL39 melanoma cell lines (>58.1% reduction, p < 0.0332), as well as the invasive capacity of the A375 melanoma cell line (>50% reduction, p < 0.0021). Expression of myeloid cell leukemia-1 (MCL1), a miR-1469 target gene, was reduced in the A375 and MEL39 cell lines by immunoblot. No significant differences in viability, resistance to apoptotic stimuli, or proliferation were observed following transfection. These findings together demonstrate how migration and invasion are specific functions through which miR-1469 expression in melanoma cells can contribute to the differences in disease progression associated with tumor ulceration.
Asunto(s)
Melanoma/genética , MicroARNs/metabolismo , Proteína 1 de la Secuencia de Leucemia de Células Mieloides/genética , Neoplasias Cutáneas/genética , Úlcera Cutánea/genética , Apoptosis/efectos de los fármacos , Apoptosis/genética , Biopsia , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Movimiento Celular/genética , Progresión de la Enfermedad , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Melanoma/patología , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , Invasividad Neoplásica/genética , Invasividad Neoplásica/prevención & control , Piel/patología , Neoplasias Cutáneas/patología , Úlcera Cutánea/patologíaRESUMEN
The polarization of macrophages plays a critical role in the pathophysiology of rheumatoid arthritis. The macrophages can have pro-inflammatory M1 polarization and various types of alternative anti-inflammatory M2 polarization. Our preliminary results showed that the CDKN2B-AS1/MIR497/TXNIP axis might regulate macrophages of rheumatoid arthritis patients. Therefore, we hypothesized that this axis regulated the polarization of rheumatoid macrophages. Flow cytometry was used to determine the surface polarization markers in M1 or M2 macrophages from healthy donors and rheumatoid arthritis patients. The QPCR and Western Blotting were used to compare the expression of the CDKN2B-AS1/MIR497/TXNIP axis in these macrophages. We Knocked down and overexpressed the axis in the macrophage cell line MD to test its roles in macrophage polarization. Compared to cells from healthy donors, cells from rheumatoid arthritis patients expressed higher levels of CD40 and CD80 and lower levels of CD16, CD163, CD206, and CD200R after polarization, they also expressed higher CDKN2B-AS1, lower MIR497, and higher TXNIP. In macrophages from healthy donors, there was no correlation among CDKN2B-AS1, MIR497, and TXNIP. But in macrophages from patients, there were significant correlations. The CDKN2B-AS1 knockdown, MIR497 mimics suppressed the M1 polarization but promoted the M2 polarization in MD cells, while the MIR497 knockdown and the TXNIP overexpression did the opposite. This study demonstrated that elevated CDKN2B-AS1 in macrophages promotes the M1 polarization and inhibited the M2 polarization of macrophages by the CDKN2B-AS1/ MIR497/TXNIP axis.
Asunto(s)
Artritis Reumatoide/inmunología , Proteínas Portadoras/genética , Macrófagos/inmunología , MicroARNs/metabolismo , ARN Largo no Codificante/metabolismo , Adulto , Anciano , Artritis Reumatoide/sangre , Artritis Reumatoide/genética , Estudios de Casos y Controles , Separación Celular , Células Cultivadas , Femenino , Citometría de Flujo , Técnicas de Silenciamiento del Gen , Voluntarios Sanos , Humanos , Activación de Macrófagos/efectos de los fármacos , Activación de Macrófagos/genética , Macrófagos/metabolismo , Masculino , MicroARNs/agonistas , Persona de Mediana Edad , Cultivo Primario de Células , ARN Largo no Codificante/genéticaRESUMEN
Cisplatin is a highly effective and broad-spectrum anticancer drug for the clinical treatment of solid tumors. However, it causes acute kidney injury (AKI) in patients with cancer. Consequently, its clinical application is limited. The occurrence, development, and prognosis of AKI are closely associated with microRNA (miRNA), which needs validation as a biomarker, especially for the early stages of cisplatin-induced AKI. An example of miRNA is miR-132-3p, which plays important roles in inflammatory responses, cell proliferation, and apoptosis in a variety of diseases. However, variations in its expression, potential mechanisms, and downstream targets in cisplatin-induced AKI remain unclear. This study aimed to investigate the functions of miR-132-3p in cisplatin-induced AKI. Sequencing and qRT-PCR revealed that miR-132-3p was significantly upregulated in cisplatin-induced AKI models of mouse and human proximal renal tubular epithelial (HK-2) cells. Apoptosis and inflammatory responses were significantly suppressed by the inhibition of the miR-132-3p expression in cisplatin-stimulated HK-2 cells, and this suppression was blocked by miR-132-3p mimics. Bioinformatics and dual luciferase reporter gene assay identified the 3'- UTR of SIRT1 mRNA as a direct target of miR-132-3p. RNA-FISH and immunofluorescence co-localization demonstrated that miR-132-3p and SIRT1 directly combined and interacted in the cytoplasm of HK-2 cells. Mechanistically, the SIRT1 expression was suppressed and the NF-κB signaling pathway was activated by the upregulation of miR-132-3p in cisplatin-induced AKI. By contrast, the SIRT1 expression was upregulated after the inhibition of miR-132-3p. The ratios of p-p65/p65 and p-IκBα/IκBα were significantly reduced, and the expression levels of inflammatory biomarkers and apoptotic proteins induced by cisplatin were obviously attenuated. Our results suggested that miR-132-3p exacerbated cisplatin-induced AKI by negatively regulating SIRT1 and activating the NF-κB signaling pathway. Therefore, targeting miR-132-3p might be a potential adjuvant therapy for ameliorating AKI in cisplatin-treated patients.
Asunto(s)
Lesión Renal Aguda/genética , Cisplatino/efectos adversos , Epigénesis Genética/efectos de los fármacos , MicroARNs/metabolismo , Sirtuina 1/genética , Acetilación , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/inmunología , Lesión Renal Aguda/patología , Animales , Apoptosis/efectos de los fármacos , Apoptosis/genética , Línea Celular , Proliferación Celular/efectos de los fármacos , Proliferación Celular/genética , Modelos Animales de Enfermedad , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/inmunología , Epigénesis Genética/inmunología , Células Epiteliales , Humanos , Túbulos Renales/efectos de los fármacos , Túbulos Renales/inmunología , Túbulos Renales/patología , Masculino , Ratones , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , FN-kappa B/metabolismoRESUMEN
Ischemia/reperfusion (I/R)induced liver injury remains a primary concern in liver transplantation and hepatectomy. Previous studies have indicated that microRNAs (miRs) are involved in multiple pathophysiological processes, including liver I/R. miR1405p reportedly inhibits inflammatory responses and apoptosis in several diseases; however, the role of miR1405p in liver I/R remains unknown. The present study aimed to investigate the potential role and mechanism of miR1405p on liver I/R injury. Mouse liver I/R and mouse AML12 cell hypoxia/reoxygenation (H/R) models were established. miR1405p mimics, inhibitor or agonists were used to overexpress or inhibit miR1405p in vitro and in vivo. Reverse transcriptionquantitative polymerase chain reaction was used to detect miR1405p expression. Liver and cell injury were evaluated using several biochemical assays. The association between miR1405p and calpain1 (CAPN1) was confirmed using a dualluciferase reporter assay. The results revealed that miR1405p expression was decreased in the mouse model of liver I/R injury and AML12 cells subjected to H/R, while overexpressed miR1405p reduced liver injury in vivo and cell injury in vitro. In addition, CAPN1 was determined to be a target of miR1405p; overexpressed CAPN1 abrogated the effect of miR1405p on H/Rinduced cell injury. The present study indicated that miR1405p protected against liver I/R by targeting CAPN1, which may provide a novel therapeutic target for liver I/R injury.
Asunto(s)
Calpaína/metabolismo , Hígado/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Daño por Reperfusión/genética , Daño por Reperfusión/metabolismo , Alanina Transaminasa/sangre , Animales , Apoptosis/genética , Calpaína/genética , Caspasa 3/metabolismo , Línea Celular , Supervivencia Celular/genética , Citocinas/metabolismo , Modelos Animales de Enfermedad , L-Lactato Deshidrogenasa/metabolismo , Hígado/lesiones , Hígado/patología , Masculino , Ratones Endogámicos C57BL , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Daño por Reperfusión/patología , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Inflammation and oxidative stress have indispensable roles in the development of acute lung injury (ALI). MicroRNA (miRNA/miR)3515p was initially identified as a myogenesisassociated miRNA; however, its role in lipopolysaccharide (LPS)induced ALI remains unclear. The aim of the present study was to investigate the role and potential mechanisms of miR3515p in ALI. ALI was induced through a single intratracheal injection of LPS for 12 h, and miR3515p agomir, antagomir or their corresponding negative controls were injected into the tail vein before LPS stimulation. Compound C, 2',5'dideoxyadenosine and H89 were used to inhibit AMPactivated protein kinase (AMPK), adenylate cyclase and protein kinase A (PKA), respectively. miR3515p levels in the lungs were significantly increased in response to LPS injection. miR3515p antagomir alleviated, while miR3515p agomir aggravated LPSinduced oxidative stress and inflammation in the lungs. The present results also demonstrated that miR3515p antagomir attenuated LPSinduced ALI via activating AMPK, and that the cAMP/PKA axis was required for the activation of AMPK by the miR3515p antagomir. In conclusion, the present study indicated that miR3515p aggravated LPSinduced ALI via inhibiting AMPK, suggesting that targeting miR3515p may help to develop efficient therapeutic approaches for treating ALI.
Asunto(s)
Proteínas Quinasas Activadas por AMP/antagonistas & inhibidores , Lesión Pulmonar Aguda/genética , Lesión Pulmonar Aguda/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Lesión Pulmonar Aguda/inducido químicamente , Animales , AMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Modelos Animales de Enfermedad , Inflamación/genética , Lipopolisacáridos/toxicidad , Masculino , Ratones Endogámicos C57BL , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Estrés Oxidativo/genéticaRESUMEN
Colorectal cancer (CRC) has become the third most common cause of cancer-related deaths. CRC occurs because of abnormal growth of cells that can invade other tissues and cause distant metastases. Researchers have suggested that aberrant microRNA (miRNA) expression is involved in the initiation and progression of cancers. However, the key miRNAs that regulate the growth and metastasis of CRC remain unclear. The circulating miRNAs from BALB/c mice with CRC CT26 cell implantation were assayed by microarray. Then, Mus musculus (house mouse) mmu-miR-762 mimic and inhibitor were transfected to CT26 cells for analysis of cell viability, invasion, and epithelial-mesenchymal transition (EMT), cell cycle, and regulatory molecule expression. Human subjects were included for comparison the circulating Homo sapiens (human) has-miR-762 levels in CRC patients and control donors, as well as the patients with and without distant metastasis. The result for miRNA levels in mice with CRC cell implantation indicated that plasma mmu-miR-762 was upregulated. Transfection of mmu-miR-762 mimic to CT26 cells increased cell viability, invasion, and EMT, whereas transfection of mmu-miR-762 inhibitor decreased the above abilities. Cells treated with high-concentration mmu-miR-762 inhibitor induced cell cycle arrest at G0/G1 phase. However, mmu-miR-762 did not cause apoptosis of cells. Western blot analysis showed that mmu-miR-762 mimic transfection upregulated the expression of Wnt-1 and ß-catenin, as well as increased the nuclear translocation of ß-catenin. Further analysis was performed to demonstrate the correlation of miR-762 with CRC, and blood samples were collected from CRC patients and control donors. The results showed that serum has-miR-762 levels in CRC patients were higher than in control donors. Among the CRC patients (n= 20), six patients with distant metastasis showed higher serum has-miR-762 levels than patients without distant metastasis. Conclusions, the present study suggests that circulating miR-762 might be a potential biomarker for upregulation of CRC cell growth and invasion, and may be accompanied by the Wnt/ß-catenin signaling.
Asunto(s)
MicroARN Circulante/metabolismo , Neoplasias Colorrectales/diagnóstico , MicroARNs/metabolismo , Animales , Línea Celular Tumoral , MicroARN Circulante/agonistas , MicroARN Circulante/antagonistas & inhibidores , MicroARN Circulante/sangre , Neoplasias Colorrectales/sangre , Neoplasias Colorrectales/genética , Modelos Animales de Enfermedad , Regulación Neoplásica de la Expresión Génica , Humanos , Masculino , Ratones , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , MicroARNs/sangre , Regulación hacia Arriba , Vía de Señalización Wnt/genéticaRESUMEN
Atherosclerosis (AS) is a dynamic and multi-stage process that involves various cells types, such as vascular smooth muscle cells (VSMCs) and molecules such as microRNAs. In this study, we investigated how miR-338-3p works in the process of AS. To determine how miR-338-3p was expressed in AS, an AS rat model was established and primary rat VSMCs were cultured. Real-time polymerase chain reaction was performed to detect miR-338-3p expression. Markers of different VSMC phenotypes were tested by Western blot. Immunofluorescent staining was employed to observe the morphologic changes of VSMCs transfected with miR-338-3p mimics. A dual luciferase reporter assay system was used to verify that desmin was a target of miR-338-3p. To further identify the role of miR-338-3p in the development of AS, VSMC proliferation and migration were evaluated by EdU incorporation assay, MTT assay, and wound healing assay. miR-338-3p expression was upregulated in the aortic tissues of an AS rat model and in primary rat VSMCs from a later passage. The transfection of miR-338-3p mimics in VSMCs promoted the synthetic cell phenotype. Bioinformatics analysis proposed desmin as a candidate target for miR-338-3p and the dual luciferase reporter assay confirmed in vivo that desmin was a direct target of miR-338-3p. The MTT and EdU incorporation assay revealed increased cell viability when miR-338-3p mimics were transfected. The increased expression of PCNA was a consistent observation, although a positive result was not obtained with respect to VSMC mobility. In AS, miR-338-3p expression was elevated. Elevated miR-338-3p inhibited the expression of desmin, thus promoting the contractile-to-synthetic VSMC phenotypic transition. In addition to morphologic changes, miR-338-3p enhanced the proliferative but not mobile ability of VSMCs. In summary, miR-338-3p promotes the development of AS.
Asunto(s)
Aorta/metabolismo , Apolipoproteínas E/genética , Aterosclerosis/genética , Desmina/genética , MicroARNs/genética , Músculo Liso Vascular/metabolismo , Animales , Aorta/patología , Apolipoproteínas E/deficiencia , Aterosclerosis/etiología , Aterosclerosis/metabolismo , Aterosclerosis/patología , Emparejamiento Base , Secuencia de Bases , Línea Celular , Movimiento Celular , Proliferación Celular , Desmina/metabolismo , Dieta Alta en Grasa/efectos adversos , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Células HEK293 , Humanos , MicroARNs/agonistas , MicroARNs/metabolismo , Músculo Liso Vascular/patología , Miocitos del Músculo Liso/metabolismo , Miocitos del Músculo Liso/patología , Oligorribonucleótidos/genética , Oligorribonucleótidos/metabolismo , Cultivo Primario de Células , Antígeno Nuclear de Célula en Proliferación/genética , Antígeno Nuclear de Célula en Proliferación/metabolismo , Ratas , Ratas Sprague-Dawley , Ratas Transgénicas , Transducción de SeñalRESUMEN
MiR-29a belongs to one of the subtypes of miRNAs known as non-coding single-stranded RNAs and is preferentially expressed in normal tissues. B7-H3, a member of the B7/CD28 immunoglobulin superfamily, was shown to be overexpressed in several solid malignant tumors, including colon cancer. In addition, it is associated with tumor progression and poor prognosis. We used immunohistochemical and Western blotting to assess B7-H3 protein expression levels in colon cancer and adjacent normal tissues and then compared their relationships with clinicopathological factors. Quantitative real-time reverse-transcription PCR was used to assess B7-H3 and miRNA-29a mRNA expression levels, and then their relationship and clinical significance were evaluated. In addition, colon cancer Caco-2 cells, which constitutively overexpress B7-H3, were transfected with lentivirus particles for miR-29a upregulation. Invasion and migration assays were carried out in vitro along with the establishment of a subcutaneous xenograft model in vivo to determine the role of miRNA-29a in colon cancer progression. The B7-H3 protein showed elevated expression in colon carcinoma and was relevant to TNM staging, lymph node metastasis, and reduced survival. Meanwhile, miR-29a was preferentially expressed in normal colon tissues, while B7-H3 transcript levels had no marked differences between tumor and normal tissue specimens. In vitro, miR-29a upregulation resulted in reduced B7-H3 expression. Furthermore, miR-29a upregulation reduced the invasive and migratory abilities of colon carcinoma cells. In animal models, upregulation of miR-29a slowed down the growth of subcutaneous xenotransplanted tumors and resulted in prolonged survival time. MiR-29a downregulates B7-H3 expression and accordingly inhibits colon cancer progression, invasion, and migration, indicating miR-29a and B7-H3 might represent novel molecular targets for advanced immunotherapy in colon cancer.
Asunto(s)
Antígenos B7/genética , Neoplasias del Colon/genética , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , Anciano , Anciano de 80 o más Años , Animales , Antígenos B7/metabolismo , Células CACO-2 , Movimiento Celular , Proliferación Celular , Neoplasias del Colon/metabolismo , Neoplasias del Colon/mortalidad , Neoplasias del Colon/patología , Progresión de la Enfermedad , Femenino , Humanos , Metástasis Linfática , Masculino , Ratones Endogámicos BALB C , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , MicroARNs/metabolismo , Persona de Mediana Edad , Estadificación de Neoplasias , Oligorribonucleótidos/genética , Oligorribonucleótidos/metabolismo , Transducción de Señal , Análisis de Supervivencia , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Multiple sclerosis (MS) is a neurodegenerative and demyelinating autoimmune disease mediated by autoreactive T cells that affects the central nervous system (CNS). Electroacupuncture (EA) has emerged as an alternative or supplemental treatment for MS, but the mechanism by which EA may alleviate MS symptoms is unresolved. Here, we examined the effects of EA at the Zusanli (ST36) acupoint on mice with experimental autoimmune encephalomyelitis (EAE), the predominant animal model of MS. The effects of EA on EAE emergence, inflammatory cell levels, proinflammatory cytokines, and spinal cord pathology were examined. EA treatment attenuated the EAE clinical score and associated spinal cord demyelination, while reducing the presence of proinflammatory cytokines in mononuclear cells (MNCs), downregulating microRNA (miR)-155, and upregulating the opioid peptide precursor proopiomelanocortin (POMC) in the CNS. Experiments in which cultured neurons were transfected with a miR-155 mimic or a miR-155 inhibitor further showed that the direct modulation of miR-155 levels could regulate POMC levels in neurons. In conclusion, the alleviation of EAE by EA is characterized by reduced proportions of Th1/Th17 cells and increased proportions of Th2 cells, POMC upregulation, and miR-155 downregulation, while miR-155 itself can suppress POMC expression. These results, support the hypothesis that the effects of EA on EAE may involve the downregulation of miR-155.
Asunto(s)
Electroacupuntura , Encefalomielitis Autoinmune Experimental/terapia , MicroARNs/metabolismo , Esclerosis Múltiple/terapia , Animales , Regulación hacia Abajo/inmunología , Encefalomielitis Autoinmune Experimental/inmunología , Femenino , Humanos , Ratones , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , Esclerosis Múltiple/inmunología , Proopiomelanocortina/genética , Células TH1/inmunología , Células Th17/inmunología , Células Th2/inmunología , Regulación hacia Arriba/inmunologíaRESUMEN
Understanding the underlying mechanisms of pediatric osteosarcoma (OS) migration and invasion is important for prognosis and treatment. We tried to measure the expression of long non-coding RNA HLA complex group 18 (HCG18) in OS and reveal its function in the malignant behaviors of OS cells. This study detected the expression of HCG18, miR-188-5p and forkhead box C1 (FOXC1) in OS tissues and cell lines by quantitative real-time PCR (qRT-PCR). The relevance between miR-188-5p and HCG18 or FOXC1 was affirmed by dual-luciferase reporter (DLR) assay. Cell viability was analyzed by MTT assay. Transwell assay was utilized to test cell invasion and migration. FOXC1 protein expression was detected by western blot. HCG18 expression was elevated in OS tissues, and enhanced HCG18 expression was related to metastasis. HCG18 silencing repressed the viability, migration and invasion of OS cells. Moreover, HCG18 interacted with miR-188-5p. MiR-188-5p up-regulation repressed cell viability, invasion and migration in OS cells. FOXC1, a known target of miR-188-5p, was negatively modulated by miR-188-5p. Furthermore, miR-188-5p inhibition or FOXC1 over-expression partially abolished the reduced of cell viability, invasion and migration mediated by HCG18 silencing in OS cell lines. This study revealed that HCG18 knockdown repressed the viability, invasion and migration of OS cells by targeting miR-188-5p and regulating FOXC1 expression. Thus, HCG18/ miR-188-5p/FOX may be a hopeful target for OS therapy.
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Neoplasias Óseas/genética , Factores de Transcripción Forkhead/genética , MicroARNs/genética , Osteoblastos/metabolismo , Osteosarcoma/genética , ARN Largo no Codificante/genética , Adolescente , Emparejamiento Base , Secuencia de Bases , Neoplasias Óseas/metabolismo , Neoplasias Óseas/patología , Línea Celular , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Supervivencia Celular , Niño , Preescolar , Femenino , Factores de Transcripción Forkhead/metabolismo , Regulación Neoplásica de la Expresión Génica , Humanos , Masculino , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , MicroARNs/metabolismo , Invasividad Neoplásica , Oligorribonucleótidos/genética , Oligorribonucleótidos/metabolismo , Osteoblastos/patología , Osteosarcoma/metabolismo , Osteosarcoma/patología , ARN Largo no Codificante/agonistas , ARN Largo no Codificante/antagonistas & inhibidores , ARN Largo no Codificante/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Transducción de Señal , Adulto JovenRESUMEN
ABSTRACT: MicroRNAs play important roles in gestational diabetes mellitus (GDM), and this study aimed to elucidate the clinical significance of miR-96-5p in diagnosing GDM.There are 123 pregnant women diagnosed with GDM and 123 healthy pregnant women were enrolled as control participants. Placenta and plasma samples from the patients and control participants were collected, and quantitative reverse transcription polymerase chain reaction (RT-qPCR) was performed to determine miR-96-5p expression levels. Moreover, a receiver operating characteristic (ROC) curve was established to evaluate the significance of miR-96-5p in diagnosing GDM. HRT-8/SVneo trophoblasts were cultured under high glucose conditions and treated with miR-96-5p mimics, and cell viability was examined.miR-96-5p levels were significantly decreased in both the placenta and plasma samples of patients with GDM. The ROC curve indicated that miR-96-5p can serve as a diagnostic biomarker for GDM with high sensitivity and specificity. Moreover, miR-96-5p levels were markedly low under high glucose conditions, and the overexpression of miR-96-5p increased the viability, respectively, of trophoblasts in vitro.miR-96-5p may participate in the pathogenesis of GDM by exerting effects on the viability of trophoblasts.
Asunto(s)
Diabetes Gestacional/diagnóstico , MicroARNs/análisis , Adulto , Biomarcadores/análisis , Biomarcadores/metabolismo , Glucemia/análisis , Estudios de Casos y Controles , Línea Celular , Supervivencia Celular/efectos de los fármacos , Medios de Cultivo/metabolismo , Diabetes Gestacional/sangre , Diabetes Gestacional/metabolismo , Diabetes Gestacional/patología , Femenino , Glucosa/metabolismo , Prueba de Tolerancia a la Glucosa , Humanos , MicroARNs/agonistas , MicroARNs/metabolismo , Placenta/patología , Embarazo , Curva ROC , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , TrofoblastosRESUMEN
BACKGROUND AND AIMS: Primary biliary cholangitis (PBC) is an autoimmune cholestatic liver disease. We found microRNA-34a (miR-34a), as the downstream gene of p53, was overexpressed in some of fibrogenic diseases. In this study, we sought to explore whether miR-34a plays a role in the fibrosis of PBC. METHODS: The peripheral blood of PBC patients and controls was collected to analyze the level of miR-34a. Human intrahepatic biliary epithelial cells (HIBEC) were cultured. The expression of miR-34a was regulated by miR-34a mimics and inhibitor. The biomarkers of epithelium-mesenchymal transition (EMT), fibrogenesis, inflammation, and transforming growth factor- (TGF-) ß1/smad pathway were analyzed. RESULTS: We found that miR-34a was overexpressed in the peripheral blood in PBC patients. In vitro, overexpressed miR-34a increased the EMT and fibrogenesis activity of HIBEC. Transforming growth factor-beta type 1 receptor (TßR1), TGF-ß1, and p-smad2/3 were upregulated by miR-34a. Inflammatory factors such as IL-6 and IL-17 were also upregulated. Finally, we showed that miR-34a promoted EMT and liver fibrosis in PBC by targeting the TGF-ß1/smad pathway antagonist transforming growth factor-beta-induced factor homeobox 2 (TGIF2). CONCLUSIONS: Our findings show that miR-34a plays an important role in the EMT and fibrosis of PBC through the TGF-ß1/smad pathway by targeting TGIF2. This study suggests that miR-34a may be a new marker of fibrogenesis in PBC. Inhibition of miR-34a may be a promising strategy in treating PBC and improving the prognosis of the disease.
Asunto(s)
Transición Epitelial-Mesenquimal/genética , Cirrosis Hepática Biliar/complicaciones , Cirrosis Hepática/genética , MicroARNs/metabolismo , Conductos Biliares/citología , Conductos Biliares/patología , Estudios de Casos y Controles , Células Epiteliales , Transición Epitelial-Mesenquimal/efectos de los fármacos , Células HEK293 , Humanos , Hígado/inmunología , Hígado/patología , Cirrosis Hepática/sangre , Cirrosis Hepática/inmunología , Cirrosis Hepática/patología , Cirrosis Hepática Biliar/sangre , Cirrosis Hepática Biliar/genética , Cirrosis Hepática Biliar/inmunología , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , Cultivo Primario de Células , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Transducción de Señal/inmunología , Proteínas Smad/metabolismo , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
Mechanical unloading-induced bone loss is a clinical challenge, and deep understanding for this disease is necessary for developing novel and effective therapies. MicroRNAs (miRNAs) are small non-coding RNAs, and involved in bone remodeling. In the study, we attempted to explore the potential of miR-133a in regulating osteoblast activation and its anti-osteopenia function both in vitro and in vivo. Our in vitro studies at first showed that miR-133a could significantly promote the expression of osteocalcin (OCN), Collagen I, alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2) and osterix (Osx), promoting the activation and mineralization of osteoblasts. Then, hindlimb unloading (HU)-challenged mice were established with or without intravenous injection of agomir-miR-133a using an osteoblast-targeting delivery system. We found that miR-133a in osteoblasts significantly alleviated the bone loss, microstructural, and biomechanical property in mice with mechanical unloading, contributing to osteopenia alleviation. Furthermore, both in vitro and in vivo experiments showed that miR-133a could restrain osteoclastogenesis via tartrate-resistant acid phosphatase (TRAP) staining. In conclusion, our results suggested that miR-133a may be a promising factor in mediating the occurrence and progression of osteopenia caused by mechanical unloading, and thus targeting miR-133a could be considered as an effective therapeutic strategy for the suppression of pathological osteopenia.
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Enfermedades Óseas Metabólicas/genética , MicroARNs/metabolismo , Osteoblastos/patología , Animales , Enfermedades Óseas Metabólicas/diagnóstico , Enfermedades Óseas Metabólicas/patología , Línea Celular , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Fémur/patología , Humanos , Masculino , Ratones , MicroARNs/agonistas , Osteoblastos/metabolismo , Osteogénesis/genética , Microtomografía por Rayos XRESUMEN
MicroRNAs (miRNAs) play a critical role in regulating the response of animals exposed to heavy metal stress. As a globally dispersed heavy metal in aquatic ecosystems, cadmium (Cd) is highly toxic to many aquatic species. However, little is known about the miRNA response to Cd stress in fish. To investigate the regulatory effect of miRNAs in response to Cd, common carp (Cyprinus carpio) were exposed to Cd2+-containing water (0.005 mg/L, 0.05 mg/L, 0.5 mg/L) for 30 days. After exposure, Cd2+ contents were significantly higher in the kidneys of C. carpio compared to other tissues, when exposed to 0.5 mg/L Cd2+. Hematoxylin and eosin staining images revealed that elevated Cd induced inflammatory damage in the kidneys of C.carpio. Further, miRNA sequencing revealed nine differentially expressed miRNAs (miR-217, miR-205 and seven novel miRNAs) in the kidneys, between 0.5 mg/L Cd2+ exposure and control groups. Potential target mRNAs of miRNAs suggest that miR-217 is involved in immunotoxicity. miR-217 agomir was intraperitoneally administered to C. carpio and RT-PCR revealed that the expression of IL-8 and SIRT1 decreased, while TLR-4, TRAF6, NF-kB, TNF-α, IL-1ß, and TGF-ß increased in the kidneys of C.carpio. Additionally, the expression of SIRT1 decreased, while the expression of other mRNAs increased in kidneys of C. carpio exposed to Cd. According to mRNAs expression in the agomir and Cd treatment, miRNAs inhibit the expressions of target mRNAs. These results demonstrate that miR-217 via SIRT1 plays a regulatory role in the immunotoxicity of Cd to C. carpio.
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Cadmio/toxicidad , Carpas , MicroARNs/metabolismo , Sirtuina 1/metabolismo , Animales , Citocinas/genética , Citocinas/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Riñón/efectos de los fármacos , Riñón/metabolismo , MicroARNs/agonistas , MicroARNs/genética , Sirtuina 1/genética , Contaminantes Químicos del Agua/toxicidadRESUMEN
Hepatocellular carcinoma is a common type of liver cancer. Resistance to chemotherapeutic agents is a major problem in cancer therapy. MicroRNAs have been reported in cancer development and tumor growth; however, the relationship between chemoresistance and hepatocellular carcinoma needs to be fully investigated. Here, we treated hepatocellular carcinoma cell line (HA22T) with a histone deacetylase inhibitor to establish hepatocellular carcinoma-resistant cells (HDACi-R) and investigated the molecular mechanisms of chemoresistance in HCC cells. Although histone deacetylase inhibitor could not enhance cell death in HDACi-R but upregulation of miR-107 decreased cell viability both in parental cells and resistance cells, decreased the expression of cofilin-1, enhanced ROS-induced cell apoptosis, and dose-dependently sensitized HDACi-R to HDACi. Further, miR-107 upregulation resulted in tumor cell disorganization in both HA22T and HDACi-R in a mice xenograft model. Our findings demonstrated that miR-107 downregulation leads to hepatocellular carcinoma cell resistance in HDACi via a cofilin-1-dependent molecular mechanism and ROS accumulation.
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Carcinoma Hepatocelular/tratamiento farmacológico , Resistencia a Antineoplásicos/genética , Inhibidores de Histona Desacetilasas/farmacología , Neoplasias Hepáticas/tratamiento farmacológico , MicroARNs/metabolismo , Animales , Apoptosis/efectos de los fármacos , Apoptosis/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Proliferación Celular/genética , Cofilina 1/genética , Resistencia a Antineoplásicos/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Técnicas de Silenciamiento del Gen , Inhibidores de Histona Desacetilasas/uso terapéutico , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patología , Masculino , Ratones , MicroARNs/agonistas , MicroARNs/genética , Especies Reactivas de Oxígeno/metabolismo , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Cerebral ischemia-reperfusion (CI/R) injury is a serious complication when treating patients experiencing ischemic stroke. Although the microRNA miR-27a-3p reportedly participates in ischemia/reperfusion (I/R) injury, its actions in CI/R remain unclear. To mimic CI/R in vitro, HT22 cells were subjected to oxygen glucose deprivation/reoxygenation (OGD/R). The results indicate that OGD inhibited growth and induced apoptosis among HT22 cells. The apoptosis was accompanied by increases in activated caspases 3 and 9 and decreases in Bcl-2. Oxidative stress was also increased, as indicated by increases in ROS and malondialdehyde and decreases in glutathione and superoxide dismutase. In addition, OGD induced G1 arrest in HT22 cells with corresponding upregulation of FOXO1 and p27 Kip1, suggesting the cell cycle arrest was mediated by FOXO1/p27 Kip1 signaling. Notably, FOXO1 was found to be the direct target of miR-27a-3p in HT22 cells. MiR-27a-3p was downregulated in OGD/R-treated HT22 cells, and miR-27a-3p mimics partially or entirely reversed all of the in vitro effects of OGD. Moreover, miR-27a-3p agomir significantly alleviated the symptoms of CI/R in vivo in a rat model of CI/R. Thus, MiR-27a-3p appears to suppress CI/R injury by targeting FOXO1.
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Proteína Forkhead Box O1/genética , Accidente Cerebrovascular Isquémico/complicaciones , MicroARNs/metabolismo , Proteínas del Tejido Nervioso/genética , Daño por Reperfusión/genética , Animales , Apoptosis/genética , Encéfalo/citología , Encéfalo/patología , Puntos de Control del Ciclo Celular/genética , Línea Celular , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Modelos Animales de Enfermedad , Regulación hacia Abajo , Proteína Forkhead Box O1/metabolismo , Humanos , Accidente Cerebrovascular Isquémico/genética , Accidente Cerebrovascular Isquémico/patología , Ratones , MicroARNs/agonistas , Proteínas del Tejido Nervioso/metabolismo , Neuronas/patología , Estrés Oxidativo/genética , Ratas , Ratas Wistar , Daño por Reperfusión/patología , Regulación hacia ArribaRESUMEN
BACKGROUNDS: A major side effect of statin, a widely used drug to treat hyperlipidemia, is skeletal myopathy through cell apoptosis. The aim of this study is to investigate the roles of microRNA in statin-induced injury. METHODS: Apolipoprotein E knockout (ApoE-/-) mice were administered with simvastatin (20 mg/kg/day) for 8 weeks. Exercise capacity was evaluated by hanging grid test, forelimb grip strength, and running tolerance test. RESULTS: In cultured skeletal muscle cells, statin increased the levels of miR-1a but decreased the levels of mitogen-activated protein kinase kinase kinase 1 (MAP3K1) in a time or dose dependent manner. Both computational target-scan analysis and luciferase gene reporter assay indicated that MAP3K1 is the target gene of miR-1a. Statin induced cell apoptosis of skeletal muscle cells, but abolished by downregulating of miR-1a or upregulation of MAP3K1. Further, the effects of miR-1a inhibition on statin-induced cell apoptosis were ablated by MAP3K1 siRNA. In ApoE-/- mice, statin induced cell apoptosis of skeletal muscle cells and decreased exercise capacity in mice infected with vector, but not in mice with lentivirus-mediated miR-1a gene silence. CONCLUSION: Statin causes skeletal injury through induction of miR-1a excessive expression to decrease MAP3K1 gene expression.
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Inhibidores de Hidroximetilglutaril-CoA Reductasas/efectos adversos , Quinasa 1 de Quinasa de Quinasa MAP/genética , MicroARNs/metabolismo , Fibras Musculares Esqueléticas/patología , Enfermedades Musculares/inducido químicamente , Animales , Apoptosis/efectos de los fármacos , Apoptosis/genética , Células Cultivadas , Modelos Animales de Enfermedad , Humanos , Hiperlipidemias/tratamiento farmacológico , Ratones , Ratones Noqueados para ApoE , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , Fibras Musculares Esqueléticas/efectos de los fármacos , Enfermedades Musculares/diagnóstico , Enfermedades Musculares/genética , Enfermedades Musculares/patología , Condicionamiento Físico Animal , Cultivo Primario de Células , ARN Interferente Pequeño/metabolismo , Simvastatina/efectos adversos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
This study was designed to investigate the role of miR-671-5p in in vitro and in vivo models of ischemic stroke (IS). Middle cerebral artery occlusion and reperfusion (MCAO/R) in C57BL/6 mice as well as oxygen-glucose deprivation and reoxygenation (OGD/R) in a mouse hippocampal HT22 neuron line were used as in vivo and in vitro models of IS injury, respectively. miR-671-5p agomir, miR-671-5p antagomir, pcDNA3.1-NF-κB, and negative controls were transfected into cells using riboFECT CP reagent. miR-671-5p agomir, pcDNA3.1-NF-κB, and negative vectors were administered into MCAO/R mice via intracerebroventricular injection. The results showed that miR-671-5p was significantly downregulated and that miR-671-5p agomir alleviated injury and neuroinflammation induced by ischemic reperfusion. A dual-luciferase reporter assay confirmed that NF-κB is a direct target of miR-671-5p. Reverse experiments showed that miR-671-5p agomir reduced neuroinflammation via suppression of NF-κB expression in both in vitro and in vivo models of IS. Our data suggest that miR-671-5p may be a viable therapeutic target for diminishing neuroinflammation in patients with IS.