RESUMEN
BACKGROUND: It has been known that chronic rhinosinusitis with nasal polyps (CRSwNP) is a type 2 inflammation-dominated disease; however, the reasons causing such type of mucosal inflammation in CRSwNP are not well elucidated. OBJECTIVE: We sought to investigate the role of microRNA-21-5p (miR-21-5p) in regulating mucosal type 2 inflammation in CRSwNP. METHODS: miR-21-5p expression was detected in nasal mucosa of patients with CRSwNP. Correlations between miR-21-5p and indicators of type 2 inflammation were further analyzed. miR-21 knockout mice were used to explore the role of miR-21-5p in a murine model of eosinophilic (E) CRSwNP. Target gene of miR-21-5p related to type 2 inflammation in CRSwNP was identified. RESULTS: The upregulated miR-21-5p in the nasal mucosa of patients with CRSwNP, compared with control subjects, was expressed higher in patients with ECRSwNP than in patients with nonECRSwNP. miR-21-5p expression was positively correlated with mucosal eosinophil infiltrations and the expression of type 2 inflammatory cytokines. In the CRSwNP mice, miR-21 knockout significantly attenuated type 2 inflammation, as indicated by eosinophil infiltrations and expression of cytokines/chemokines in nasal mucosa and lavage fluid; moreover, genes associated with type 2 inflammation were extensively downregulated at the transcriptome level in miR-21 knockout mice. Glucagon-like peptide-1 receptor, which was negatively correlated with miR-21-5p expression in human nasal mucosa, was identified as the target of miR-21-5p. Overexpression of miR-21-5p induced IL-33 expression, whereas glucagon-like peptide-1 receptor agonist decreased IL-33 production in airway epithelial cells. CONCLUSIONS: miR-21-5p aggravates type 2 inflammation in the nasal mucosa of patients with CRSwNP via targeting glucagon-like peptide-1 receptor/IL-33 signaling, which may be a potential therapeutic target for CRSwNP.
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MicroARNs , Pólipos Nasales , Humanos , Ratones , Animales , Pólipos Nasales/genética , Interleucina-33/genética , Receptor del Péptido 1 Similar al Glucagón , Ratones Noqueados , MicroARNs/genéticaRESUMEN
Our previous findings revealed that hsa_circ_0068,888 was markedly down-regulated in the plasma of patients with sepsis-associated acute kidney injury (AKI). However, its molecular mechanism in AKI remains unclear. Herein, we explored the role of hsa_circ_0068,888 in AKI. Human renal proximal tubular cell line HK-2 was stimulated with lipopolysaccharide (LPS) to mimic AKI in vitro. Decreased hsa_circ_0068,888 expression was observed in AKI cell model. The overexpression of hsa_circ_0068,888 significantly increased the viability of LPS-stimulated HK-2 cells, whereas hsa_circ_0068,888 downregulation showed the opposite effect. Furthermore, LPS triggered inflammatory response and oxidative stress, which was inhibited by hsa_circ_0068,888 overexpression and enhanced by hsa_circ_0068,888 down-regulation. Hsa_circ_0068,888 overexpression suppressed the activation of nuclear factor-κB (NF-κB) pathway triggered by LPS as evidenced by decreased p-p65 protein level and nuclear translocation of p65 in hsa_circ_0068,888 overexpressed cells. Additionally, we proved that hsa_circ_0068,888 targeted microRNA-21-5p (miR-21-5p). The expression of miR-21-5p was markedly increased and was negatively regulated by hsa_circ_0068,888 in LPS-stimulated HK-2 cells. Furthermore, we demonstrated that miR-21-5p overexpression reversed the effects on cell viability, inflammatory response, oxidative stress, and NF-κB pathway induced by hsa_circ_0068,888 overexpression in LPS-stimulated HK-2 cells. Overall, these results implied that hsa_circ_0068,888 shows a protective effect on AKI by sponging miR-21-5p. Hence, up-regulation of hsa_circ_0068,888 might be a potential strategy in treatment for AKI.
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Túbulos Renales Proximales/metabolismo , Lipopolisacáridos/inmunología , MicroARNs/genética , ARN Circular/genética , Lesión Renal Aguda/genética , Lesión Renal Aguda/patología , Secuencia de Bases , Línea Celular , Supervivencia Celular/genética , Humanos , Inflamación/genética , Túbulos Renales Proximales/citología , Túbulos Renales Proximales/inmunología , Túbulos Renales Proximales/patología , FN-kappa B/metabolismo , Estrés Oxidativo/genéticaRESUMEN
Erectile dysfunction (ED) is a common comorbidity in males with diabetes mellitus (DM), whose pathogenesis might be induced by dysregulation of corpus cavernosum smooth muscle cells (CCSMCs). Gene Expression Omnibus repository-based analysis identified the differentially expressed PDCD4 in DM rats. PDCD4 has also been determined as a putative gene under the regulatory control of microRNA-21-5p (miR-21-5p). This study aimed to further determine the functional role of miR-21-5p in CCSMCs in a rat model of diabetes mellitus-induced erectile dysfunction (DMED). CCSMCs were isolated from penile cavernous tissue and cultured in high glucose (HG) medium. The expression of miR-21-5p and/or PDCD4 was altered in CCSMCs, as directly or indirectly measured by CCK-8 assay, flow cytometry, and TUNEL assays. Furthermore, exosomes were isolated from mesenchymal stem cells (MSCs) transfected with miR-21-5p mimic or miR-21-5p inhibitor and co-cultured with CCSMCs. DMED rats were injected with lentivirus carrying PDCD4/siRNA-PDCD4 plasmids, or exosomes from MSCs containing miR-21-5p-agomir to explore their roles in vivo. The experimental data validated that PDCD4 was upregulated in cavernous tissue of DMED rats. miR-21-5p targeted and inhibited PDCD4. miR-21-5p was enriched in MSC-exosomes. Moreover, PDCD4 downregulation, miR-21-5p elevation or MSC-derived exosomal miR-21-5p reduced apoptosis and enhanced proliferation of CCSMCs cultured in HG medium. PDCD4 silencing or miR-21-5p-containing MSC-exosomes improved erectile function and smooth muscle density in DMED rats. Collectively, our findings suggested that MSC-derived exosomal miR-21-5p suppressed PDCD4 expression and ED in rats with DM.
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Proteínas Reguladoras de la Apoptosis/genética , Diabetes Mellitus Experimental/complicaciones , Disfunción Eréctil/metabolismo , Exosomas/genética , MicroARNs/metabolismo , Miocitos del Músculo Liso/metabolismo , Proteínas de Unión al ARN/genética , Animales , Proteínas Reguladoras de la Apoptosis/metabolismo , Células Cultivadas , Regulación hacia Abajo , Disfunción Eréctil/etiología , Disfunción Eréctil/genética , Disfunción Eréctil/terapia , Exosomas/metabolismo , Exosomas/trasplante , Masculino , Trasplante de Células Madre Mesenquimatosas/métodos , MicroARNs/genética , Miocitos del Músculo Liso/fisiología , Pene/citología , Pene/metabolismo , Pene/fisiopatología , Proteínas de Unión al ARN/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
This study aimed to investigate the potential effects of miR-215, with exosomes as carriers, against skeletal muscle injury. Exosomes were isolated from rat bone marrow mesenchymal stem cells (rBMSCs) or rBMSCs overexpressing miR-215. Subsequently, rat myoblasts (L6) were treated with different exosomes and mimics, then exposed to H2O2. Cell viability and apoptosis were determined using the Cell Counting Kit-8 and Annexin V-FITC cell apoptosis assay kits, respectively. Reverse-transcriptase quantitative PCR (RT-qPCR) was used to examine the expression of related genes. Transmission electron microscopy, Nanosight, and western blotting showed that the exosomes were successfully isolated. PKH67 staining revealed that both exosomes and miR-215-modified exosomes were taken up by L6 cells. FABP3 was found to be the target gene of miR-215 via a dual luciferase reporter gene assay. In the L6 cells treated with H2O2, cell viability was significantly inhibited, whereas apoptosis significantly increased (P < 0.05). Exosomes significantly enhanced the viability of H2O2-induced cells and inhibited their apoptosis (P < 0.05). In addition, RT-qPCR showed that in the H2O2-induced L6 cells, FABP3, CDKN1A, and TP53 were significantly upregulated, while CCNB1 was significantly downregulated (P < 0.05). However, their expression levels were significantly reversed after treatment with miR-215-modified exosomes (P < 0.05). These findings indicate that the miR-215-modified exosomes may exert protective effects against skeletal muscle injury through the miR-215/FABP3 pathway and regulate the expression of CDKN1A, CCNB1, and TP53.
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Citoprotección , Exosomas/metabolismo , Proteína 3 de Unión a Ácidos Grasos/metabolismo , Peróxido de Hidrógeno/toxicidad , Células Madre Mesenquimatosas/metabolismo , MicroARNs/metabolismo , Mioblastos/metabolismo , Regiones no Traducidas 3'/genética , Animales , Apoptosis/efectos de los fármacos , Apoptosis/genética , Secuencia de Bases , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/genética , Citoprotección/efectos de los fármacos , Exosomas/efectos de los fármacos , Exosomas/ultraestructura , Proteína 3 de Unión a Ácidos Grasos/genética , Regulación de la Expresión Génica/efectos de los fármacos , Células Madre Mesenquimatosas/efectos de los fármacos , MicroARNs/genética , Mioblastos/efectos de los fármacos , Ratas Sprague-DawleyRESUMEN
OBJECTIVES: To explore the molecular mechanism for thyroid cancer metastasis via analyzing the role of microRNA (miR)-21-5p and its target gene recombinant sclerostin domain containing protein 1 (SOSTDC1) in thyroid cancer. METHODS: The target miR-21-5p was screened through bioinformatics analysis and cell verification, and the thyroid cancer cell lines was transfected with miR-21-5p inhibitor. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) test, flow cytometry, and cell scratch test were used to detect the proliferation, apoptosis and migration of thyroid cancer cells in the miR-21-5p inhibitor group and the inhibitor control group, respectively. The luciferase report experiment was used to verify the relationship between miR-21-5p and SOSTDC1, Western blotting was used to detect the expression levels and phosphorylation levels of SOSTDC1,phosphatidylinositol 3 kinase (PI3K), protein kinase B (Akt) and mitogen-activated protein kinases (MAPK), extracellular regulated protein kinases (ERK) in thyroid cancer cells. RESULTS: MiR-21-5p was significantly increased in thyroid cancer cells,which was negatively correlated with SOSTDC1 (r=-0.24, P<0.01). The proliferation and migration of thyroid cancer cells in the miR-21-5p inhibitor group was significantly lower than that in the inhibitor control group (both P<0.01), and the apoptosis rate in the miR-21-5p inhibitor group was significantly higher than that in the inhibitor control group (P<0.01).The luciferase report experiment showed that miR-21-5p could target and regulate the expression level of SOSTDC1, and the expression of PI3K in the miR-21-5p inhibitor group was significantly lower than that in the inhibitor control group (P<0.01). There were no significant changes in Akt and ERK1/2 levels, but the phosphorylation levels of Akt and ERK1/2 in the miR-21-5p inhibitor group were significantly lower than those in the inhibitor control group (both P<0.01). CONCLUSIONS: MiR-21-5p in thyroid cancer cells can target the expression of SOSTDC1 and affect the activities of PI3K/Akt and MAPK/ERK, thereby inhibiting the apoptosis of thyroid cancer cells and promoting cell proliferation and migration.
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MicroARNs , Neoplasias de la Tiroides , Proteínas Adaptadoras Transductoras de Señales , Apoptosis/genética , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Humanos , MicroARNs/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Neoplasias de la Tiroides/genéticaRESUMEN
Fibroblast-myofibroblast transdifferentiation (FMT) is widely recognized as the major pathological feature of renal fibrosis. Although melatonin has exerted antifibrogenic activity in many diseases, its role in renal FMT remains unclear. In the present study, the aim was to explore the effect of melatonin on renal FMT and the underlying mechanisms. We established the transforming growth factor (TGF)-ß1 stimulated rat renal fibroblast cells (NRK-49F) model in vitro and unilateral ureteral obstruction (UUO) mice model in vivo. We assessed levels of α-smooth muscle actin (α-SMA), col1a1 and fibronectin, STAT3 and AP-1, as well as miR-21-5p and its target genes (Spry1, PTEN, Smurf2 and PDCD4). We found that melatonin reduced the expression of α-SMA, col1a1 and fibronectin, as well as the formation of α-SMA filament in TGF-ß1-treated NRK-49F cells. Meanwhile, melatonin inhibited STAT3 phosphorylation, down-regulated miR-21-5p expression, and up-regulated Spry1 and PTEN expression. Moreover, miR-21-5p mimics partially antagonized the anti-fibrotic effect of melatonin. For animal experiments, the results revealed that melatonin remarkably ameliorated UUO-induced renal fibrosis, attenuated the expression of miR-21-5p and pro-fibrotic proteins and elevated Spry1 and PTEN expression. Nevertheless, agomir of miR-21-5p blocked the renoprotective effect of melatonin in UUO mice. These results indicated that melatonin could alleviate TGF-ß1-induced renal FMT and UUO-induced renal fibrosis through down-regulation of miR-21-5p. Regulation of miR-21-5p/PTEN and/or miR-21-5p/Spry1 signal might be involved in the anti-fibrotic effect of melatonin in the kidneys of UUO mice.
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Transdiferenciación Celular/efectos de los fármacos , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Enfermedades Renales/etiología , Melatonina/farmacología , Miofibroblastos/citología , Miofibroblastos/efectos de los fármacos , Actinas/genética , Actinas/metabolismo , Animales , Biomarcadores , Supervivencia Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Susceptibilidad a Enfermedades , Fibroblastos/metabolismo , Fibrosis , Expresión Génica , Humanos , Enfermedades Renales/metabolismo , Enfermedades Renales/patología , Masculino , Ratones , Ratones Noqueados , Miofibroblastos/metabolismo , Fosfohidrolasa PTEN/metabolismo , Ratas , Factor de Crecimiento Transformador beta1/farmacologíaRESUMEN
Circular RNA YAP1 (circYAP1) was reported to participate in progression of gastric cancer. However, the role of circYAP1 in acute kidney injury (AKI) remains obscure. We attempted to examine the effects of circYAP1 on ischaemia/reperfusion-stimulated renal injury. AKI model was established by treating HK-2 cells in ischaemia/reperfusion (I/R) environment. CircYAP1 expression in blood of AKI patients and I/R-treated HK-2 cells was evaluated via RT-qPCR. CCK-8, flow cytometry, ELISA and ROS assay were executed to test the impact of circYAP1 on cell viability, apoptosis, inflammatory cytokines and ROS generation. Bioinformatic analysis was executed to explore miRNA targets. The relativity between circYAP1 and miR-21-5p was verified by RT-qPCR and luciferase assay. The functions of miR-21-5p in I/R-triggered injury were reassessed. PI3K/AKT/mTOR pathway was detected by Western blot. Down-regulated circYAP1 was observed in AKI blood samples and I/R-treated HK-2 cells. CircYAP1 overexpression expedited cell growth and weakened secretion of inflammatory factors and ROS generation in I/R-disposed cells. Besides, we found circYAP1 could sponge to miR-21-5p. Interestingly, miR-21-5p overexpression overturned the repressive effects of circYAP1 on cell injury. Moreover, PI3K/AKT/mTOR pathway was activated by circYAP1 via inhibiting miR-21-5p. We demonstrated that circYAP1 activated PI3K/AKT/mTOR pathway and secured HK-2 cells from I/R injury via sponging miR-21-5p.
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Lesión Renal Aguda/genética , Proteínas Adaptadoras Transductoras de Señales/genética , MicroARNs/genética , ARN Circular/genética , Daño por Reperfusión/genética , Factores de Transcripción/genética , Lesión Renal Aguda/patología , Apoptosis , Proliferación Celular/genética , Supervivencia Celular/genética , Regulación de la Expresión Génica/genética , Humanos , Fosfatidilinositol 3-Quinasas/genética , Proteínas Proto-Oncogénicas c-akt/genética , Daño por Reperfusión/patología , Transducción de Señal/genética , Serina-Treonina Quinasas TOR/genética , Proteínas Señalizadoras YAPRESUMEN
The patho-mechanism leading to airway wall remodeling in allergic asthma is not well understood and remodeling is resistant to therapies. This study assessed the effect of immunoglobulin E (IgE) in the absence of allergens on human primary airway smooth muscle cell (ASMC) remodeling in vitro. ASMCs were obtained from five allergic asthma patients and five controls. Proliferation was determined by direct cell counts, mitochondrial activity by expression of cytochrome c, protein expression by immunoblotting and immuno-fluorescence, cell migration by microscopy imaging, and collagen deposition by cell based ELISA and RNA expression by real time PCR. Non-immune IgE activated two signaling pathways: (i) signal transducer and activator of transcription 3 (STAT3)âmiR-21-5pâdownregulating phosphatase and tensin homolog (PTEN) expression, and (ii) phosphatidylinositol 3-kinases (PI3K)âprotein kinase B (Akt)âmammalian target of rapamycin (mTOR)âribosomal protein S6 kinase beta-1 (p70s6k)âperoxisome proliferator-activated receptor gamma coactivator 1-α (PGC1-α)âperoxisome proliferator-activated receptor-γ (PPAR-γ)âcyclooxygenase-2 (COX-2)âmitochondrial activity, proliferation, migration, and extracellular matrix deposition. Reduced PTEN expression correlated with enhanced PI3K signaling, which upregulated ASMC remodeling. The inhibition of microRNA-21-5p increased PTEN and reduced mTOR signaling and remodeling. Mimics of microRNA-21-5p had opposing effects. IgE induced ASMC remodeling was significantly reduced by inhibition of mTOR or STAT3. In conclusion, non-immune IgE alone is sufficient for stimulated ASMC remodeling by upregulating microRNA-21-5p. Our findings suggest that the suppression of micoRNA-21-5p may present a therapeutic target to reduce airway wall remodeling.
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Remodelación de las Vías Aéreas (Respiratorias) , Regulación hacia Abajo , Inmunoglobulina E/metabolismo , MicroARNs/metabolismo , Miocitos del Músculo Liso/metabolismo , Fosfohidrolasa PTEN/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Asma/inmunología , Asma/patología , Movimiento Celular , Matriz Extracelular/metabolismo , Femenino , Humanos , Masculino , MicroARNs/genética , Persona de Mediana Edad , Mitocondrias/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptores de IgE/metabolismo , Proteínas Quinasas S6 Ribosómicas 70-kDa/metabolismo , Factor de Transcripción STAT3/metabolismo , Transducción de Señal , Regulación hacia ArribaRESUMEN
BACKGROUND: This study aimed to explore the effects of microRNA-21-5p (miR-21-5p) on the radiation sensitivity of non-small cell lung cancer (NSCLC) and the involvement of human MutS homolog 2 (hMSH2) One hundred fourteen NSCLC patients at stage II or III who received surgery and postoperative radiotherapy were enrolled in this study. METHODS: The patients were assigned into radiation-sensitive and -insensitive groups. NSCLC A549 cells were transfected to generate control, Negative control (NC), miR-21-5p inhibitor, miR-21-5p mimic, small interfering hMSH2 (sihMSH2), miR-21-5p inhibitor + sihMSH2 and hMSH2 overexpression groups. Immunohistochemistry was performed to detect the hMSH2 expression in transfected and irradiated cells. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were performed to evaluate A549 miR-21-5p and hMSH2 expression in transfected and irradiated cells. A colony formation assay was adopted for cell survival analysis. The relationship between miR-21-5p and hMSH2 was verified by a luciferase reporter assay. Cell viability was measured by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, and apoptosis was assessed by flow cytometry. NSCLC nude mouse models were established, and tumor volumes and tumor weights were recorded. RESULTS: The radiation-sensitive group of patients exhibited lower miR-21-5p but higher hMSH2 expression than the insensitive group. For irradiated A549 cells, lower cell survival, higher apoptosis, increased miR-21-5p expression and decreased hMSH2 expression were observed at 6 and 8 Gy than at 0, 2 and 4 Gy; compared to 6 Gy, cell survival and hMSH2 expression were decreased and apoptosis and miR-21-5p expression were increased at 8 Gy. Additionally, miR-21-5p was found to target hMSH2. Compared with the control group, the cell survival rate was lower and the apoptosis rate higher in the miR-21-5p inhibitor group, whereas the opposite was observed for the miR-21-5p mimic and sihMSH2 groups. For the mouse model, decreased tumor volume and tumor weight and higher hMSH2 expression were found in the miR-21-5p inhibitor, radiation, hMSH2 overexpression, miR-21-5p inhibitor + radiation and hMSH2 overexpression + radiation groups compared with the control group. In addition, tumor volume and tumor weight were decreased and hMSH2 expression increased in the miR-21-5p inhibitor + radiation and hMSH2 overexpression + radiation groups compared with the radiation alone group. CONCLUSION: These findings indicate that inhibition of miR-21 can promote the radiation sensitivity of NSCLC by targeting hMSH2.
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Apoptosis/efectos de la radiación , Carcinoma de Pulmón de Células no Pequeñas/patología , Rayos gamma , Neoplasias Pulmonares/patología , MicroARNs/metabolismo , Proteína 2 Homóloga a MutS/metabolismo , Células A549 , Anciano , Animales , Antagomirs/metabolismo , Secuencia de Bases , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/radioterapia , Femenino , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/radioterapia , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , MicroARNs/antagonistas & inhibidores , MicroARNs/genética , Persona de Mediana Edad , Proteína 2 Homóloga a MutS/antagonistas & inhibidores , Proteína 2 Homóloga a MutS/genética , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Tolerancia a Radiación , Alineación de Secuencia , Trasplante HeterólogoRESUMEN
Colorectal cancer is one of the most common malignant tumors worldwide with rising incidence. MicroRNAs are small non-coding RNAs that implicate in multiple physiological or pathological processes. The aberrant expression of miRNA-215 (miR-215) has been illustrated in various types of cancers. However, the expression of miR-215 in human colon cancer and the biological roles of it remain largely unknown. We conducted this study to explore the expression and the function of miR-215 in human colon cancer. The results showed that miR-215 was remarkably downregulated in colon cancer tissues and cell lines. Overexpression of miR-215 by miR-215 mimic significantly inhibited colon cancer cell proliferation, migration and invasion while knockdown of miR-215 by miR-215 inhibitor exerted reverse effects. Furthermore, we newly identified Yin-Yang 1(YY1) as a direct target of miR-215 which could rescue the effects of miR-215 on colon cancer cells. In summary, our investigation revealed that miR-215 was downregulated in colon cancer and it suppressed colon cancer cell proliferation, migration and invasion by directly targeting YY1.
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Neoplasias del Colon/metabolismo , Regulación Neoplásica de la Expresión Génica , MicroARNs/metabolismo , Factor de Transcripción YY1/metabolismo , Apoptosis , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Neoplasias del Colon/patología , Regulación hacia Abajo , Células HCT116 , Humanos , Invasividad NeoplásicaRESUMEN
Rheumatoid arthritis (RA) is characterized by progressive joint destruction with subsequent serious disability. Objective biomarkers of RA course progression are lacking, which necessitates the discovery of activity indicators and predictors of the disease outcome. Musculoskeletal Ultrasound Seven-joint Score (MSUS7) is proposed as a reliable technique to evaluate radiographic RA progression. Homo sapiens-microRNA-21-5p (hsa-miR-21-5p) plays an important role during joint remodeling and the pro-inflammatory process driving RA progression. We aimed to evaluate plasma hsa-miR-21-5p as a noninvasive RA activity biomarker and to investigate if hsa-miR-21-5p is linked to MSUS7 components in the context of RA activity. This cross-sectional study included 71 RA patients classified into inactive (n = 36) and active (n = 35) groups according to the Disease Activity Score 28-joint count with ESR (DAS28-ESR). Joints were assessed by MSUS7. Gray-scale ultrasound (GSUS) and power Doppler ultrasound (PDUS) were used to rate the synovitis, tenosynovitis, and erosion in the joints. Plasma hsa-miR-21-5p expression was measured by real-time PCR. The absolute count of regulatory T cell (Treg) was calculated after Treg frequency was assessed by flow cytometry. Results: Hsa-miR-21 expression was significantly up-regulated in the active RA group with a median fold change of 51.6 in comparison to the inactive cases with a median fold change of 7.7 (p < 0.001). Hsa-miR-21-5p was positively correlated with DAS28-ESR, C reactive protein (CRP), and rheumatoid factor (r = 0.7, p < 0.001, r = 0. 0.6, p < 0.001, and r = 0.4, p = 0.002, respectively), while negatively correlated with Treg absolute count (r = -0.4, p < 0.001). Hsa-miR-21-5p levels were correlated with synovitis and tenosynovitis in GSUS (r = 0.4, p < 0.001, r = 0.3, p = 0.025, respectively) and in PDUS (r = 0.5, p < 0.001 and 0.4, p = 0.001, respectively). The hsa-miR-21-5p accurately distinguished RA activity [AUC 0.933, 94.3% sensitivity, and 86.1% specificity]. Logistic regression analysis revealed hsa-miR-21-5p as an independent predictor for RA flare (OR = 1.228, p = 0.004). Hsa-miR-21-5p was linked to synovitis and tenosynovitis components of the MSUS7. Up-regulated hsa-miR-21-5p can be utilized as a predictor for RA disease flare.
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Artritis Reumatoide , MicroARNs , Sinovitis , Tenosinovitis , Humanos , Tenosinovitis/diagnóstico por imagen , Estudios Transversales , Brote de los Síntomas , Artritis Reumatoide/diagnóstico por imagen , Ultrasonografía , Sinovitis/diagnóstico por imagen , Biomarcadores , Índice de Severidad de la EnfermedadRESUMEN
Considering the limited research and the prevailing evidence of STAT4's tumor-suppressing role in breast carcinoma (BC) or in breast radiotherapy (RT) sensitivity requires more in-depth exploration. Our study delves into how STAT4, a transcription factor, affects BC cell resistance to radiotherapy by regulating the MALAT1/miR-21-5p/THRB axis. Bioinformatics analysis was performed to predict the regulatory mechanisms associated with STAT4 in BC. Subsequently, we identified the expression profiles of STAT4, MALAT1, miR-21-5p, and THRB in various tissues and cell lines, exploring their interactions and impact on RT resistance in BC cells. Moreover, animal models were established with X-ray irradiation for further validation. We discovered that STAT4, which is found to be minimally expressed in breast carcinoma (BC) tissues and cell lines, has been associated with a poorer prognosis. In vitro cellular assays indicated that STAT4 could mitigate radiotherapy resistance in BC cells by transcriptional activation of MALAT1. Additionally, MALAT1 up-regulated THRB expression by adsorbing miR-21-5p. As demonstrated in vitro and in vivo, overexpressing STAT4 inhibited miR-21-5p and enhanced THRB levels through transcriptional activation of MALAT1, which ultimately contributes to the reversal of radiotherapy resistance in BC cells and the suppression of tumor formation in nude mice. Collectively, STAT4 could inhibit miR-21-5p and up-regulate THRB expression through transcriptional activation of MALAT1, thereby mitigating BC cell resistance to radiotherapy and ultimately preventing BC development and progression.
RESUMEN
BACKGROUND: Adipose-derived mesenchymal stem cells (adMSCs) are suggested as potential tools for the treatment of regenerative diseases, including tissue repair. This study aimed to explore the function of adMSC-derived apoptotic bodies in skin wound healing and the molecules of action. METHODS: The acquired adMSCs and their-derived apoptotic bodies were identified. A murine model of full-thickness skin wounds was treated with apoptotic bodies. The wound healing process of mice and the pathological changes in wound tissues were examined. Ana-1 macrophages were treated with lipopolysaccharide (LPS) and apoptotic bodies for in vitro experiments. Polarization of macrophages was examined by immunofluorescence staining of the specific biomarkers and ELISA kits. Dermal microvascular endothelial cells (DMECs) or dermal fibroblasts (DFs) were co-cultured with apoptotic bodies or the LPS- and apoptotic bodies-treated Ana-1 cells. Downstream molecules mediated by apoptotic bodies were screened by microarray and bioinformatic analyses. RESULTS: Apoptotic bodies treatment accelerated skin wound healing in mice and promoted formation of granulation tissues and blood vessels in wound tissues. Apoptotic bodies treatment induced M2 polarization of macrophages. The angiogenesis ability of DMECs, and the viability and migration of DFs were increased when co-cultured with the apoptotic bodies-treated Ana-1 cells. MicroRNA (miR)-21-5p was abundantly expressed in ABs, and kruppel like factor 6 (KLF6) mRNA was confirmed as a target of miR-21-5p. Overexpression of KLF6 reduced M2 polarization of macrophages and blocked the promoting effect of apoptotic bodies on wound healing in vitro and in vivo. CONCLUSION: miR-21-5p carried by adMSC-derived apoptotic bodies targets KLF6 to induce M2 polarization of macrophages and augment skin wound healing.
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Quemaduras , Vesículas Extracelulares , Células Madre Mesenquimatosas , MicroARNs , Animales , Ratones , Factor 6 Similar a Kruppel , Células Endoteliales , Lipopolisacáridos/farmacología , Quemaduras/terapia , Cicatrización de Heridas , Macrófagos , MicroARNs/genéticaRESUMEN
Aim: Head and neck squamous cell carcinoma (HNSCC) is the sixth most commonly diagnosed malignancy worldwide. Overexpressed of microRNA-21-5p (miR-21-5p) has been reported to be involved in the development of HNSCC. However, the role of miR-21-5p in HNSCC is still not fully elucidated. The purpose of this study was to explore the underlying molecular mechanisms of miR-21-5p in HNSCC. Methods: RT-qPCR was used to determine the differential expression levels of miR-21-5p in tissue samples of HNSCC patients. Meta-analysis was performed based on miRNA expression data collected from the Gene Expression Omnibus (GEO) database, The Cancer Genome Atlas (TCGA), and published articles to evaluate the expression of miR-21-5p in HNSCC. We investigated the biological function of miR-21-5P by gene ontology enrichment and target prediction analysis. Furthermore, RT-qPCR and IHC were conducted to verify the expression of target genes. Finally, Kaplan-Meier survival analysis was performed to assessed the prognostic value of the putative miR-21-5p target genes. Results: MiR-21-5p was significantly overexpressed in HNSCC compared to healthy tissues (P < .05) and showed potent predictive power with a summary receiver operating characteristic of 0.90. Meanwhile, the expression of miR-21-5p was significantly correlated with tumor stage, T stage and smoking in HNSCC (P < .05). A total of 71 down-regulated genes, both HNSCC-related and miR-21-p5-related, were obtained from the analytical integration. Two predicted genes (ADH7, RDH12) were down-regulated in HNSCC, and significantly negatively correlated with miR-21-5p. IHC and RT-qPCR demonstrated that the expression of ADH7 and RDH12 in HNSCC samples was significantly lower than control. And high expression of ADH7 was associated with better DFS of HNSCC patients. Conclusions: miR-21-5p may target at ADH7, RDH12 and participate in regulation of retinol metabolism, which might affect the prognosis of HNSCC. High expression of ADH7 may indicate better prognosis in HNSCC patients.
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Neoplasias de Cabeza y Cuello , MicroARNs , Oxidorreductasas de Alcohol/genética , Oxidorreductasas de Alcohol/metabolismo , Biomarcadores de Tumor/genética , Biología Computacional , Regulación Neoplásica de la Expresión Génica , Neoplasias de Cabeza y Cuello/genética , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello/genéticaRESUMEN
Background: Cardiovascular disease, including acute myocardial infarction (AMI), is a major global cause of mortality and morbidity. Specificity and sensitivity limit the utility of classic diagnostic biomarkers for AMI. Therefore, it is critical to identify novel biomarkers for its accurate diagnosis. Cumulative studies have demonstrated that circulating microRNAs (miRs) participate in the pathophysiological processes of AMI and are promising diagnostic biomarkers for the condition. This study aimed to ascertain the diagnostic accuracy of circulating miR-21-5p and miR-126 used as biomarkers in patients with AMI and infarct-related artery total occlusion (IR-ATO) or infarct-related blood-vessel recanalization (IR-BVR). Methods: The expression of miR-21-5p and miR-126 was examined separately in 50 healthy subjects, 51 patients with IR-ATO AMI, and 49 patients with IR-BVR AMI using quantitative real-time polymerase chain reaction. Results: When compared with the control group, the IR-ATO AMI group exhibited increased miR-21-5p (p < 0.0001) and miR-126 (p < 0.0001), and the IR-BVR AMI group exhibited increased miR-21-5p (p < 0.0001). However, there was no significant difference in miR-126 between the IR-BVR AMI and the control groups. A Spearman's correlation coefficient showed a strong correlation was found between miR-21-5p, miR-126, cardiac troponin-I, and creatine kinase isoenzyme in all three groups, while a receiver operating characteristic analysis revealed that miR-21-5p and miR-126 exhibited considerable diagnostic accuracy for IR-ATO AMI. Conclusion: Circulating miR-21-5p and miR-126 may be promising prognostic biomarkers for patients with AMI and IR-ATO.
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Long non-coding (lnc) RNAs play crucial functions in human cancer. However, until recently, the involvement of the lncRNA GAS6-AS1 in breast cancer (BCa) malignancy has not been studied exhaustively. The roles and underlying mode of action of GAS6-AS1 action in BCa progression were examined through functional experiments. A decline in GAS6-AS1 level led to a significant decrease in BCa cell proliferation, and the ability for colony formation. Here, GAS6-AS1 competed as endogenous RNA by sequestering microRNA-215-5p (miR-215-5p) causing an enhanced expression of SRY-box transcription factor 9 (SOX9). The effects of silencing GAS6-AS1 on BCa malignant phenotypes could be ameliorated by inhibiting miR-215-5p or restoring SOX9. Thus, GAS6-AS1 acted as a lncRNA that drives tumor in BCa, and enabled progression of BCa through miR-215-5p /SOX9 axis regulation. These outcomes show that the GAS6-AS1/miR-215-5p/SOX9 axis is a potentially effective target for cancer treatment and management.
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Ropivacaine (Rop) is available to suppress the growth of glioblastoma (GBM), while its mechanism has not been completely elaborated. In this study, we explore the latent mechanism of Rop repressing GBM's growth via mediating the microRNA (miR)-21-5p/KAT8 regulatory NSL complex subunit 2 (KANSL2) axis. MiR-21-5p was declined in GBM, while KANSL2 was elevated. Clinical association studies manifested miR-21-5p was distinctly linked to the tumor size and grade of GBM. Rop constrained GBM cell proliferation, invasion, and migration but boosted apoptosis. Elevated miR-21-5p strengthened Rop's action, while augmented KANSL2 weakened Rop's role. Furthermore, the impact of silencing miR-21-5p on GBM was turned around via declining KANSL2 in Rop-treated GBM cells. KANSL2 was the target gene of miR-21-5p. In short, Rop exerted an anti-tumor impact on GBM via mediating the miR-21-5p/KANSL2 axis, which offered novel viewpoints for the later adoption of Rop as GBM drugs.
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Neoplasias Encefálicas , Glioblastoma , Histona Acetiltransferasas , MicroARNs , Ropivacaína , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patología , Línea Celular Tumoral , Proliferación Celular/genética , Glioblastoma/genética , Glioblastoma/patología , Histona Acetiltransferasas/genética , Humanos , MicroARNs/genética , Ropivacaína/farmacologíaRESUMEN
Background: This study aimed to investigate the function of circular RNA La-related protein 4 (circ-LARP4) on non-small cell lung cancer (NSCLC) progression. Materials and Methods: Circ-LARP4 overexpression and circ-LARP4 short hairpin RNA (shRNA) plasmids were transfected into NCI-H1650 cells and NCI-H1299 cells respectively. In rescue experiment, microRNA (miR)-21-5p overexpression and miR-21-5p shRNA plasmids were transfected into circ-LARP4 overexpression-treated NCI-H1650 cells and circ-LARP4 knockdown-treated NCI-H1650 cells, respectively. Circ-LARP4 and miR-21-5p expression levels were detected by reverse transcription-quantitative polymerase chain reaction. Cell proliferation and apoptosis were investigated by cell counting kit-8 assay and annexin V/propidium iodide assay. The interaction between circ-LARP4 and miR-21-5p was further explored by luciferase reporter assay. Results: Circ-LARP4 expression was decreased in NSCLC cell lines (including NCI-H1299, NCI-H522, NCI-H23, NCI-H358, and NCI-H1650) compared with human normal lung epithelial cell line. Circ-LARP4 overexpression inhibited cell proliferation while promoted apoptosis in NCI-H1650 cells, whereas circ-LARP4 knockdown increased cell proliferation while decreased apoptosis in NCI-H1299 cells. Meanwhile, miR-21-5p was negatively regulated by circ-LARP4, whereas circ-LARP4 was not affected by miR-21-5p in NCI-H1650 and NCI-H1299 cells. In rescue experiment, miR-21-5p overexpression attenuated the effect of circ-LARP4 overexpression on decreasing cell proliferation and increasing apoptosis in NCI-H1650 cells, whereas miR-21-5p knockdown attenuated the effect of circ-LARP4 knockdown on promoting cell proliferation and suppressing apoptosis in NCI-H1299 cells. Further luciferase reporter assay revealed that circ-LARP4 could directly bind to miR-21-5p. Conclusions: Circ-LARP4 is decreased and suppresses cell proliferation while promoted apoptosis by sponging miR-21-5p in NSCLC.
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Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , MicroARNs , Apoptosis/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , Línea Celular Tumoral , Proliferación Celular/genética , Humanos , Neoplasias Pulmonares/genética , MicroARNs/genética , MicroARNs/metabolismo , ARN Circular/genéticaRESUMEN
BACKGROUND AND STUDY AIMS: Recent reports have emphasized the increased risk of hepatocellular carcinoma (HCC) post direct-acting antiviral (DAAs) therapy in chronic hepatitis C virus (HCV) patients. Unfortunately, reliable diagnostic markers for HCC are still lacking. In this context, serum microRNAs have become promising diagnostic targets. Thus, the current study aims to elaborate the diagnostic utility of microRNA 122-5p, microRNA 21-5p, and microRNA 222-3p in the serum of Egyptian patients presenting with HCV infection and HCC post DAA therapy. PATIENTS AND METHODS: Qiagen specific microRNA assays were utilized to assess the expression levels of the chosen microRNAs in the serum samples collected from 3 groups: (1) 50 patients with HCV-related HCC, (2) 50 patients with HCC post DAA therapy, and 20 healthy control. RESULTS: The mean serum values of microRNA 21-5p and microRNA 122-5p were significantly lower in the HCC post DAA therapy group than in both the group with HCC without prior exposure to DAAs (P < 0.001) and control group (P 0.05 and 0.02, respectively). A significant upregulation was observed for both microRNA 21-5p and microRNA 122-5p in the HCV-related HCC group compared with the control group (P < 0.001 and = 0.011, respectively). On the other hand, the mean serum value of microRNA 222-3p was significantly raised in the HCC post DAA therapy group than in the control group (P = 0.007), whereas no statistically significant difference was observed between both groups with HCC and between the group with HCV-related HCC without prior exposure to DAAs and control group. CONCLUSION: This is the first study to introduce microRNA 21-5p, microRNA 122-5p and microRNA 222-3p as noninvasive biomarker candidates for HCC post DAA therapy. Their altered expression among treatment-naive HCC and HCC post DAA therapy might assume a different microRNA profiling in both HCC groups.
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Carcinoma Hepatocelular , Hepatitis C Crónica , Neoplasias Hepáticas , MicroARNs , Antivirales/uso terapéutico , Carcinoma Hepatocelular/virología , Hepatitis C Crónica/complicaciones , Hepatitis C Crónica/tratamiento farmacológico , Hepatitis C Crónica/patología , Humanos , Neoplasias Hepáticas/virología , MicroARNs/genéticaRESUMEN
MicroRNAs (miRNAs or miRs) play important roles in cardiovascular disease. miR-21-5p is known to be involved in the regulation of cardiomyocyte injury under high glucose and high fat (HG-HF) conditions, but its mechanism of action remains unclear. In the present study, a cardiomyocyte cell line, H9c2, was treated with 33 mM glucose and 250 µM sodium palmitate for 24, 48, and 72 h to produce HG-HF injury. After treatment, miR-21-5p expression was detected by reverse transcription-quantitative PCR. A miR-21-5p mimic was then constructed and transfected into the cells and the potential molecular mechanism was investigated using Cell Counting Kit-8, TUNEL, flow cytometry and western blot assays. Expression of miR-21-5p was significantly downregulated by HG-HF treatment of H9c2 cells for 24, 48, and 72 h. In subsequent experiments, cells were treated for an intermediate period (48 h). Compared with the control group, HG-HF treatment significantly inhibited H9c2 proliferation and promoted apoptosis, while these effects were significantly reduced in the miR-21-5p mimic. Compared with the control group, HG-HF treatment significantly increased reactive oxygen species, while miR-21-5p mimic significantly reduced this effect. Compared with the control group, HG-HF treatment significantly increased the expression of the pro-apoptotic proteins Bax and phosphorylated (p)-Akt and decreased the expression of the anti-apoptotic proteins Bcl-2, p-PTEN, and p-FOXO3a, while overexpression of miR-21-5p significantly reduced these effects. The results revealed that miR-21-5p inhibited apoptosis and oxidative stress in H9c2 cells induced by HG-HF, likely through the PTEN/Akt/FOXO3a signaling pathway.