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Objective: MicroRNAs (miRNAs) are involved in a range of pathological and biological processes. Vascular involvement is an important complication associated with morbidity and mortality in Behçet's disease (BD). In this study, we aimed to evaluate the expression levels of miR-195, miR-424, miR-10b, miR-103a-3p, and miR-542-3p in Turkish patients with BD, and their possible association with vascular involvement and clinical activity. Methods: This cross-sectional study included 61 BD patients and 25 age- and sex-matched healthy individuals. The patients were categorised into two groups based on the presence or absence of vascular involvement. Demographic data, disease duration, disease activity, and medical treatments were recorded. Disease activity was evaluated using the Behçet's Disease Current Activity Form (BDCAF) and the Behçet's Syndrome Activity Scale (BSAS). The expression levels of miRNAs were measured using real-time quantitative polymerase chain reaction (RT-qPCR). Results: The comparison of the clinical features of BD patients with and without vascular involvement revealed no significant difference. However, the expression levels of miR-195, miR-424, miR-10b, miR-103a-3p, and miR-542-3p were significantly higher in BD patients than in healthy controls (p<0.001, p<0.001, p=0.010, p<0.01, p=0.039, respectively). Moreover, the expression level of miR-195 was significantly higher in vasculo-Behçet patients than in the other groups (p=0.0318). However, no significant association was found between the expression levels of miR-195 and clinical activity. Conclusion: Our study results indicated elevated serum levels of miR-195 in BD patients, which may be associated with vascular involvement. Therefore, miR-195 could potentially serve as a biomarker for the diagnosis and monitoring of vasculo-Behçet's disease.

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The complex etiologies and mechanisms of Alzheimer's disease (AD) underscore the importance for devising multitarget drugs to achieve effective therapy. MicroRNAs (miRNAs) are capable of concurrently regulating the expression of multiple proteins by selectively targeting disease- associated genes in a sequence-specific fashion. Nonetheless, as RNA-based drugs, their stability in the circulation and capacity of traversing the blood-brain barrier (BBB) is largely compromised, thereby limiting their potential clinical applications. In this study, we formulated the nanoliposomes encapsulating polyethyleneimine (PEI)/miR-195 complex (DPMT@PEI/miR-195) that was engineered through dual modifications to contain P-aminophenyl-alpha-d-mannopyranoside (MAN) and cationic cell-penetrating peptide (TAT). DPMT@PEI/miR-195 exhibited the enhanced BBB- and cell membrane penetrating capability. As expected, we observed that DPMT@PEI/miR-195 administered through intravenous tail injection of produced greater effectiveness than donepezil and the same range of effect as aducanumab in alleviating the cognitive decline in 7-month-old APP/PS1 mice. Moreover, the combination treatment with DPMT@PEI/miR-195 and donepezil effectively ameliorated the deterioration of cognition in 16-month-old APP/PS1 mice, with enhanced effects than either DPMT@PEI/miR-195 or donepezil alone. Furthermore, DPMT@PEI/miR-195 effectively attenuated the positive signals of Aß, AT8, and CD68 in APP/PS1 mice without notable side effects. Our findings indicate DPMT@PEI/miR-195 as a promising potentially new agent or approach for the prophylaxis and treatment of early and advanced stages of Alzheimer's disease.

Asunto(s)

Enfermedad de Alzheimer , MicroARNs , Humanos , Ratones , Animales , Lactante , Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/metabolismo , Liposomas/uso terapéutico , Precursor de Proteína beta-Amiloide/metabolismo , Donepezilo/uso terapéutico , Ratones Transgénicos , MicroARNs/genética , MicroARNs/uso terapéutico , MicroARNs/metabolismo , Modelos Animales de Enfermedad , Péptidos beta-Amiloides/metabolismo

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Long intergenic non-protein coding RNA 1547 (LINC01547) presents a notable relationship with prognosis in patients with ovarian cancer. Herein, we examined the expression of LINC01547 in non-small cell lung cancer (NSCLC) to ascertain its clinical significance. We also explored the detailed functions of LINC01547 in regulating the aggressive phenotype of NSCLC and the molecular mechanism of action underlying its carcinogenic activities events in NSCLC. Furthermore, we applied the data acquired from the tissue specimens and the Cancer Genome Atlas (TCGA) database to analyze the level of LINC01547 in NSCLC and conducted functional assays to address the regulatory effect of LINC01547. Further, we examined the mechanistic interaction among LINC01547, microRNA-195-5p (miR-195-5p), and homeobox C8 (HOXC8) using bioinformatics prediction and luciferase reporter assay. LINC01547 was noticeably overexpressed, as affirmed by data from TCGA and our own cohort; moreover, poor prognosis was associated with increased LINC01547 levels in patients with NSCLC. LINC01547 regulates cell proliferation, colony-forming, migration, and invasion, and its absence produced tumor-repressing effects in NSCLC. Mechanistically, as a competitive endogenous RNA, LINC01547 decoyed miR-195-5p and consequently resulted in the overexpression of HOXC8 in NSCLC cells. Using rescue experiments, we found that the regulatory activities of LINC01547 deficient in repressing the malignant properties of NSCLC cells could be counteracted by hindering miR-195-5p or overexpressing HOXC8. Conclusively, LINC01547 serves as a crucial component to worsen the oncogenicity of NSCLC cells by controlling the miR-195-5p/HOXC8 axis. Thus, the newly identified competing endogenous RNA pathway may potentially be an attractive therapeutic for NSCLC management.

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Due to their high prevalence, gastrointestinal cancers are one of the key causes of cancer-related death globally. The development of drug-resistant cancer cell populations is a major factor in the high mortality rate, and it affects about half of all cancer patients. Because of advances in our understanding of cancer molecular biology, non-coding RNAs (ncRNAs) have emerged as critical factors in the initiation and development of gastrointestinal cancers. Gene expression can be controlled in several ways by ncRNAs, including through epigenetic changes, interactions between microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) and proteins, and the function of lncRNAs as miRNA precursors or pseudogenes. As lncRNAs may be detected in the blood, circulating ncRNAs have emerged as a promising new class of non-invasive cancer biomarkers for use in the detection, staging, and prognosis of gastrointestinal cancers, as well as in the prediction of therapy efficacy. In this review, we assessed the role lncRNAs play in the progression, and maintenance of colorectal cancer, and how they might be used as therapeutic targets in the future.

Asunto(s)

Neoplasias Gastrointestinales , MicroARNs , ARN Largo no Codificante , Humanos , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , ARN no Traducido/genética , Neoplasias Gastrointestinales/genética , Epigénesis Genética

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OBJECTIVE: To analyze the expression and correlation of microRNA-195 (miR-195), miR-125 and calreticulin in diffuse large B-cell lymphoma (DLBCL). METHODS: From April 2020 to April 2021, 80 DLBCL patients with complete data archived by the Pathology Department of Handan First Hospital and The Second Hospital of Hebei Medical University were selected as the study group, and 70 patients with reactive lymph node hyperplasia were selected as the control group. The expressions of miR-195 and miR-125 were detected by real-time fluorescence quantitative PCR, and the expression of calreticulin was detected by Western blot. Pearson correlation was used to analyze the correlation between miR-195, miR-125, calreticulin and DLBCL, and ROC curve was used to analyze the predictive value of miR-195, miR-125 and calreticulin for DLBCL. RESULTS: Compared with the control group, the expression of miR-195 decreased but miR-125 and calreticulin increased in the study group (P<0.001). The expression levels of miR-195, miR-125 and calreticulin were not related to sex, age, primary site and B symptoms of patients with DLBCL, but related to immunophenotype, Ann Arbor stage, lactate dehydrogenase, IPI score, nodule involvement and Ki-67 index. The expression of miR-195 decreased and the expression of miR-125 and calreticulin increased in DLBCL paitents with non-germinal center source, Ann Arbor stage III-IV, lactate dehydrogenase > 245 U/L, IPI score 3-5, nodule involvement≥2 and Ki-67 index≥75% (P<0.05). Pearson correlation analysis showed that miR-195 and miR-125 were negatively correlated (r=-0.536, P=0.001), miR-195 and calreticulin were negatively correlated (r=-0.545, P=0.001), while miR-125 and calreticulin were positively correlated (r=0.523, P=0.001). ROC curve showed that compared with the single diagnosis of miR-195, miR-125 and calreticulin, the combination of the three items had higher predictive value for DLBCL (P<0.001). CONCLUSION: The expression of miR-195 decreases and the expression of miR-125 and calreticulin increase in patients with DLBCL. Along with the increase of disease stage and IPI score, the decrease of miR-195 and the increase of miR-125 and calreticulin aggravate gradually. The three items may participate in the occurrence and progress of DLBCL.

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Linfoma de Células B Grandes Difuso , MicroARNs , Humanos , MicroARNs/genética , Antígeno Ki-67/metabolismo , Calreticulina/metabolismo , Pronóstico , Linfoma de Células B Grandes Difuso/genética , Lactato Deshidrogenasas/metabolismo

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OBJECTIVE@#To analyze the expression and correlation of microRNA-195 (miR-195), miR-125 and calreticulin in diffuse large B-cell lymphoma (DLBCL).@*METHODS@#From April 2020 to April 2021, 80 DLBCL patients with complete data archived by the Pathology Department of Handan First Hospital and The Second Hospital of Hebei Medical University were selected as the study group, and 70 patients with reactive lymph node hyperplasia were selected as the control group. The expressions of miR-195 and miR-125 were detected by real-time fluorescence quantitative PCR, and the expression of calreticulin was detected by Western blot. Pearson correlation was used to analyze the correlation between miR-195, miR-125, calreticulin and DLBCL, and ROC curve was used to analyze the predictive value of miR-195, miR-125 and calreticulin for DLBCL.@*RESULTS@#Compared with the control group, the expression of miR-195 decreased but miR-125 and calreticulin increased in the study group (P<0.001). The expression levels of miR-195, miR-125 and calreticulin were not related to sex, age, primary site and B symptoms of patients with DLBCL, but related to immunophenotype, Ann Arbor stage, lactate dehydrogenase, IPI score, nodule involvement and Ki-67 index. The expression of miR-195 decreased and the expression of miR-125 and calreticulin increased in DLBCL paitents with non-germinal center source, Ann Arbor stage III-IV, lactate dehydrogenase > 245 U/L, IPI score 3-5, nodule involvement≥2 and Ki-67 index≥75% (P<0.05). Pearson correlation analysis showed that miR-195 and miR-125 were negatively correlated (r=-0.536, P=0.001), miR-195 and calreticulin were negatively correlated (r=-0.545, P=0.001), while miR-125 and calreticulin were positively correlated (r=0.523, P=0.001). ROC curve showed that compared with the single diagnosis of miR-195, miR-125 and calreticulin, the combination of the three items had higher predictive value for DLBCL (P<0.001).@*CONCLUSION@#The expression of miR-195 decreases and the expression of miR-125 and calreticulin increase in patients with DLBCL. Along with the increase of disease stage and IPI score, the decrease of miR-195 and the increase of miR-125 and calreticulin aggravate gradually. The three items may participate in the occurrence and progress of DLBCL.

Asunto(s)

Humanos , MicroARNs/genética , Antígeno Ki-67/metabolismo , Calreticulina/metabolismo , Pronóstico , Linfoma de Células B Grandes Difuso/genética , Lactato Deshidrogenasas/metabolismo

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Objective To investigate the expression of serum long non-coding RNA MEG3 and microRNA(miR)-195-5p in patients with severe necrotizing pancreatitis and their relationship with the severity and prognosis of severe necrotizing pancreatitis.Methods A total of 122 patients with acute pancreatitis admitted to Cangzhou Central Hospital from October 2020 to January 2023 were selected as the research objects.Ac-cording to the severity of the disease,the patients were divided into severe necrotizing pancreatitis(severe group,53 cases)and non-severe necrotizing pancreatitis(non-severe group,69 cases).According to the prog-nosis of alternate ending with severe necrotizing pancreatitis can be divided into good prognosis group(38 ca-ses)and poor prognosis group(15 cases).At the same time,50 healthy people who underwent physical exami-nation in the hospital during the same period were selected as the control group.The clinical data and serum levels of lncRNA MEG3 and miR-195-5p in each group were compared.Spearman correlation analysis was used to analyze the relationship between serum levels of lncRNA MEG3,miR-195-5p and acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)and Ranson scores.Multivariate Logistic regression was used to analyze the influencing factors of poor prognosis in patients with severe necrotizing pancreatitis.The receiver operating characteristic(ROC)curve was used to analyze the value of lncRNA MEG3 and miR-195-5p in eval-uating the prognosis of patients with severe necrotizing pancreatitis.Results There was no significant differ-ence in age,gender,body mass index,underlying disease and etiology between severe group and non-severe group(P>0.05).Compared with the non-severe group,APACHEⅡ and Ranson scores were significantly in-creased in the severe group(P<0.05).Compared with the control group,the serum levels of lncRNA MEG3 and miR-195-5p in the non-severe group and the severe group were decreased(P<0.05),and the serum levels of lncRNA MEG3 and miR-195-5p in the severe group were lower than those in the non-severe group(P<0.05).Spearman correlation analysis showed that serum levels of lncRNA MEG3 and miR-195-5p in AP pa-tients were negatively correlated with APACHEⅡ and Ranson scores(P<0.05).Multivariate Logistic re-gression analysis showed that APACHEⅡ and Ranson scores and serum levels of lncRNA MEG3 and miR-195-5p were independent risk factors for poor prognosis in patients with severe necrotizing pancreatitis(P<0.05).ROC curve results showed that the area under the curve(AUC)of lncRNA MEG3 and miR-195-5p for evaluating the poor prognosis of patients with severe necrotizing pancreatitis was 0.767 and 0.777,respectively,the sensitivity was 86.7%and 80.0%,and the specificity was 49.9%and 45.8%,respectively.The AUC of combined e-valuation was 0.982,and the sensitivity and specificity were 86.7%and 78.8%,respectively.Conclusion The serum levels of lncRNA MEG3 and miR-195-5p are related to the severity and prognosis of severe necrotizing pancreatitis,which can evaluate the severity and predict the prognosis of severe necrotizing pancreatitis.

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BACKGROUND: Epigenetics exerts a vital role in the onset and development of renal cell carcinoma (RCC). Mounting evidence has shed light on the significance of human immune system in response to tumor infiltrating T cells. Hereby, we sought to unmask the immunomodulatory role of histone deacetylase 3 (HDAC3) and its potential upstream molecule, programmed cell death 5 (PDCD5) in RCC. METHODS: RCC and adjacent non-cancerous tissues were clinically resected from 58 patients, in which the expression profile of microRNA-195-5p (miR-195-5p), PDCD5, HDAC3, and serum glucocorticoid-inducible kinase 1 (SGK1) was determined by RT-qPCR and Western blot analysis. Their relations were investigated by a series of luciferase assays in combination with ChIP and co-IP. RCC cells (A498) were intervened using gain- and loss-of-function approaches, followed by cell proliferation evaluation. After co-culture with CD3+ T cells, flow cytometry and interferon-γ (IFN-γ) determination were performed. A xenograft tumor mouse model was developed for in vivo validation. RESULTS: PDCD5 was downregulated in RCC tissues and A498 cells. Upregulation of HDAC3, as well as of SGK1, resulted in suppression of A498 cell proliferation and promotion of T cell activation as evidenced by higher IFN-γ expression. Re-expression of PDCD5 downregulated HDAC3, causing a subsequent upregulation of miR-195-5p, while miR-195-5p could inversely modulate its target gene, SGK1. The regulatory mechanism appeared to be functional in vivo. CONCLUSION: Our results highlight the possible manipulation by PDCD5 on RCC cell proliferation and T cell activation, which provides new clues to better understand the immune balance in RCC progression.

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Proteínas Reguladoras de la Apoptosis , Carcinoma de Células Renales , Neoplasias Renales , MicroARNs , Proteínas de Neoplasias , Animales , Humanos , Ratones , Proteínas Reguladoras de la Apoptosis/genética , Carcinoma de Células Renales/metabolismo , Línea Celular Tumoral , Metilación de ADN , Interferón gamma/genética , Neoplasias Renales/patología , MicroARNs/genética , MicroARNs/metabolismo , Proteínas de Neoplasias/genética , Linfocitos T/metabolismo

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Purpose: Bone marrow-derived mesenchymal stem cells (BMSCs) are hopeful in promoting bone regeneration as their pluripotency in differentiation. Our previous study showed that carbon monoxide-releasing molecule-3 (CORM-3) increased the osteogenic differentiation of rat BMSCs in vitro. However, the mechanism remained unclear. MicroRNAs (miRNAs) play a very important role in modulating the osteogenic differentiation of BMSCs. Therefore, we researched the miRNAs involved in CORM-3-stimulated osteogenic differentiation. Methods: The CORM-3-stimulated osteogenic differentiation of rat BMSCs was further studied in vivo. Based on the gene sequencing experiment, the rat BMSCs were transfected with miR-195-5p mimics and inhibitor, pcDNA3.1-Wnt3a and Wnt3a siRNA. The osteogenic differentiation of rat BMSCs was measured by quantitative real-time polymerase chain reaction, Western blot and alizarin red staining. Additionally, the targeting relationship between miR-195-5p and Wnt3a was confirmed by the dual-luciferase assay. Results: MiR-195-5p was down-expressed during the CORM-3-stimulated osteogenic differentiation of rat BMSCs. CORM-3-stimulated osteogenic differentiation of rat BMSCs was inhibited with miR-195-5p overexpression, evidenced by significantly reduced mRNA and protein expressions of runt-related transcription factor 2 and osteopontin, and matrix mineralization demonstrated. On the contrary, the osteogenic differentiation was enhanced with inhibition of miR-195-5p. CORM-3-stimulated osteogenic differentiation of rat BMSCs was increased by overexpression of Wnt3a, while the opposite was observed in the Wnt3a-deficient cells. Moreover, the decreased osteogenic differentiation capacity by increased expression of miR-195-5p was rescued by Wnt3a overexpression, showing miR-195-5p directly targeted Wnt3a. Conclusion: These results demonstrate that CORM-3 promoted osteogenic differentiation of rat BMSCs via miR-195-5p/Wnt3a, which bodes well for the application of CORM-3 in the treatment of periodontal disease and other bone-defect diseases.

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Células Madre Mesenquimatosas , MicroARNs , Animales , Células de la Médula Ósea , Monóxido de Carbono/metabolismo , Diferenciación Celular , Células Cultivadas , Células Madre Mesenquimatosas/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Osteogénesis/genética , Ratas

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Long non-coding RNA (lncRNA) small nucleolar RNA host gene 1 (SNHG1) has been found to be highly expressed in gastric cancer (GC). However, the study for exploring the effects of SNHG1 and microRNA (miR)-195-5p on GC is limited. This research commits to unravel the regulatory effects of SNHG1, miRNA-195-5p, and Yes-associated protein 1 (YAP1) on GC. SNHG1, miR-195-5p and YAP1 levels in GC tissues and GC cells were detected. The GC cells were treated with various constructs altering SNHG1 or miR-195-5p expression to determine the biological activities of GC cell in vitro. The effect of SNHG1 inhibition on subcutaneous tumorigenesis of GC cells in a nude mouse model in vivo was detected. The binding relation among SNHG1, miR-195-5p, and YAP1 was validated. SNHG1 and YAP1 levels were elevated and miR-195-5p level was reduced in GC. Reduction of SNHG1 or elevation of miR-195-5p retarded GC cell biological activity in vitro. Downregulated SNHG1 suppressed tumor growth in vivo. SNHG1 bound to miR-195-5p, and miR-195-5p directly targeted YAP1. The downregulated SNHG1 hinders the biological behaviors of GC cells via the modulation of the miR-195-5p/YAP1 axis.

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MicroARNs , ARN Largo no Codificante , Neoplasias Gástricas , Animales , Línea Celular Tumoral , Proliferación Celular/genética , Regulación Neoplásica de la Expresión Génica , Vía de Señalización Hippo , Ratones , MicroARNs/genética , MicroARNs/metabolismo , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , ARN Nucleolar Pequeño , Neoplasias Gástricas/genética , Proteínas Señalizadoras YAP

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Objective: LINC00662 is oncogenic in some human cancers, but no much was revealed concerning to its specific action in tumor angiogenesis. Given that, our study investigated the role of LINC00662 from esophageal squamous cell carcinoma (ESCC) cells-derived extracellular vehicles (EVs) in angiogenesis through microRNA (miR)-195-5p/vascular endothelial growth factor A (VEGFA) axis. Methods: Clinical tissue samples were collected from patients with ESCC, in which LINC00662, miR-195-5p and VEGFA expression was analyzed. ESCC cells were transfected, from which EVs were isolated. Human umbilical vein endothelial cells (HUVECs) were co-cultured with the pretreated EVs. After that, viability, colony formation ability, invasion, migration and tube formation ability of HUVECs were observed. Tumor xenograft in nude mice was performed to detect the effect of LINC00662, miR-195-5p or EV specific inhibitor GW4869 on tumor development. Results: LINC00662 and VEGFA were upregulated while miR-195-5p was downregulated in the cancer tissue of patients with ESCC. EVs derived from ESCC cells promoted viability, colony formation ability, invasion and tube formation ability of HUVECs. Downregulation of LINC00662 or upregulation of miR-195-5p reversed the promotion of EVs derived from ESCC cells on the viability, colony formation ability, invasion and tube formation ability of HUVECs in vitro and in vivo. VEGFA overexpression reversed EVs carrying restored miR-195-5p induced effects on HUVECs in vitro. Conclusion: In summary, elevated LINC00662 transferred by ESCC cells-derived EVs induces angiogenesis through downregulating miR-195-5p and upregulating VEGFA.

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Circulating miR-195-5p has been proposed as a promising peripheral biomarker for the diagnosis, prognosis and severity assessment of various diseases. However, the demand for its sensitive and convenient quantification has not been met yet. Herein, we proposed a one-pot isothermal approach, in which the target signal acquisition, amplification and conversion (fluorescence read-out) system was integrated by a triple strand displacement amplification (SDA) cascade. Using this triple SDA strategy, miR-195-5p can be at least detected at 1 aM, and the linear dynamic range (from 100 aM to 1 pM) is wide enough to meet the detection needs of clinical miRNA level. A proof-of-principle study, using this novel methodology to directly analyze the spiking serum samples with different levels of miR-195-5p, demonstrated the potential of circulating miR-195-5p detection for clinical point-of-care assay. This one-pot isothermal triple SDA approach, we believe, will be a simple and feasible tool for ultrasensitive quantification of circulating miR-195-5p, and may promote the wide application of this potential biomarker in non-invasive clinical diagnosis.

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MicroARNs , Bioensayo , Biomarcadores , Humanos , MicroARNs/sangre , MicroARNs/genética , Pruebas en el Punto de Atención

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Hemangioma (HA), which is characterized by aberrant endothelial cell proliferation in blood vessels, is a common tumor during infancy. MicroRNAs (miRNAs/miRs) collectively participate in the development of HA; however, the potential roles of miR-195-5p in HA are not completely understood. The aim of the present study was to investigate the roles of miR-195-5p in HA. In the present study, miR-195-5p was found to be downregulated in HA cells, such as the XPTS-1 human infantile hemangioma-derived endothelial cell line and the EOMA hemangioendothelioma cell line. Overexpression of miR-195-5p was shown to suppress HA cell viability, colony formation and proliferation, and induced HA cell apoptosis. Furthermore, miR-195-5p downregulated Bcl-2 expression and upregulated Bax and Bcl-2 expression levels. V-ski sarcoma viral oncogene homolog (SKI) was identified as a target of miR-195-5p. Co-transfection of miR-195-5p mimics and SKI 3'-untranslated region wild-type decreased HA cell luciferase activity. SKI overexpression alleviated the miR-195-5p-induced decrease in HA cell proliferation and increased HA cell apoptosis. In addition, the regulatory role of miR-195-5p on the expression of Bcl-2, Bax and poly(ADP-ribose) polymerase was reversed by SKI. Collectively, the results of the present study demonstrated that miR-195-5p suppressed HA progression and its effects were mediated via SKI. Therefore, the miR-195-5p/SKI axis may represent a novel therapeutic target for HA.

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Calcification of the bicuspid aortic valve (BAV) involves differential expression of various RNA genes, which is achieved through complex regulatory networks that are controlled in part by transcription factors and microRNAs. We previously found that miR-195-5p regulates the osteogenic differentiation of valvular interstitial cells (VICs) by targeting the TGF-ß pathway. However, the transcriptional regulation of miR-195-5p in calcified BAV patients is not yet clear. In this study, stenotic aortic valve tissues from patients with BAVs and tricuspid aortic valves (TAVs) were collected. Candidate transcription factors of miR-195-5p were predicted by bioinformatics analysis and tested in diseased valves and in male porcine VICs. SP2 gene expression and the corresponding protein levels in BAV were significantly lower than those in TAV, and a low SP2 expression level environment in VICs resulted in remarkable increases in RNA expression levels of RUNX2, BMP2, collagen 1, MMP2, and MMP9 and the corresponding proteins. ChIP assays revealed that SP2 directly bound to the transcription promoter region of miR-195-5p. Cotransfection of SP2 shRNA and a miR-195-5p mimic in porcine VICs demonstrated that SP2 repressed SMAD7 expression via miR-195-5p, while knockdown of SP2 increased the mRNA expression of SMAD7 and the corresponding protein and attenuated Smad 2/3 expression. Immunofluorescence staining of diseased valves confirmed that the functional proteins of osteogenesis differentiation, including RUNX2, BMP2, collagen 1, and osteocalcin, were overexpressed in BAVs. In Conclusion, the transcription factor Sp2 is expressed at low levels in VICs from BAV patients, which has a negative impact on miR-195-5p expression by binding its promoter region and partially promotes calcification through a SMAD-dependent pathway.

Asunto(s)

Enfermedad de la Válvula Aórtica Bicúspide/patología , Calcinosis/patología , Osteoblastos/patología , Proteína smad7/metabolismo , Factor de Transcripción Sp2/metabolismo , Factor de Crecimiento Transformador beta1/metabolismo , Válvula Tricúspide/patología , Animales , Enfermedad de la Válvula Aórtica Bicúspide/genética , Enfermedad de la Válvula Aórtica Bicúspide/metabolismo , Proteína Morfogenética Ósea 2/genética , Proteína Morfogenética Ósea 2/metabolismo , Calcinosis/genética , Calcinosis/metabolismo , Diferenciación Celular , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Femenino , Humanos , Masculino , MicroARNs , Persona de Mediana Edad , Osteoblastos/metabolismo , Osteogénesis , Proteína smad7/genética , Factor de Transcripción Sp2/genética , Porcinos , Factor de Crecimiento Transformador beta1/genética , Válvula Tricúspide/metabolismo

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BACKGROUND: Mitochondrial dysfunction is an early feature of Alzheimer's disease (AD) and miR-195 is involved in mitochondrial disorder through targeting MFN-2 protein in hippocampal neurons of AD. OBJECTIVE: To clarify if administration of miR-195 inhibitor could enhance the memory deficits through improving hippocampal neuron mitochondrial dysfunction in SAMP8 mice. METHODS: The expression of miR-195 was detected by RT-qPCR in primary hippocampal neurons and HT-22 cells treated with Aß1-42. Morris water maze (MWM) was used to assess the learning and memory function in SAMP8 mice administrated with antagomir-195. Transmission electron microscopy was employed to determine the morphological changes of synapses and mitochondria of hippocampus in SAMP8 mice. Mitochondrial respiration was measured using a high-resolution oxygraph. RESULTS: The expression of miR-195 were upregulated in the primary hippocampal neurons and HT-22 cells induced by Aß1-42. Inhibition of miR-195 ameliorated the mitochondrial dysfunction in HT-22 cells induced by Aß1-42, including mitochondrial morphologic damages, mitochondrial membrane potential, respiration function, and ATP production. Administration of antagomir-195 by the third ventricle injection markedly ameliorated the cognitive function, postsynaptic density thickness, length of synaptic active area, mitochondrial aspect ratio, and area in hippocampus of SAMP8 mice. Finally, antagomir-195 was able to promote an increase in the activity of respiratory chain complex CI and II in SAMP8 mice. CONCLUSION: This study demonstrated that miR-195 inhibitor ameliorated the cognitive impairment of AD mice by improving mitochondrial structure damages and dysfunction in the hippocampal neurons, which provide an experimental basis for further exploring the treatment strategy of AD.

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Memoria/efectos de los fármacos , MicroARNs/efectos de los fármacos , Neuronas/efectos de los fármacos , Sinapsis/efectos de los fármacos , Enfermedad de Alzheimer/metabolismo , Animales , Apoptosis , Disfunción Cognitiva/metabolismo , Modelos Animales de Enfermedad , Femenino , GTP Fosfohidrolasas , Hipocampo/metabolismo , Masculino , Ratones , Mitocondrias/metabolismo

RESUMEN

BACKGROUND: Long noncoding RNA urothelial cancer-associated 1 (lnc-UCA1) targets microRNA-26a (miR-26a) and microRNA-195 (miR-195) to participate in coronary heart disease (CHD) progression via regulation of vascular smooth muscle cell and microvascular endothelial cell viability and mobility. Therefore, this study set out to further explore the relationship between lnc-UCA1 and miR-26a and miR-195, along with their roles in the management of patients with CHD. METHODS: One hundred and thirty-six CHD patients and 70 age-/gender-matched controls were recruited in this case-control study. Their peripheral blood mononuclear cell samples were collected for lnc-UCA1, miR-26a, and miR-195 measurement. Furthermore, serum samples from CHD patients were obtained for inflammatory cytokines and cell adhesion molecules measurement. The Gensini score was used to evaluate the stenosis severity in CHD patients. RESULTS: Lnc-UCA1 expression tend to be increased, while miR-26a and miR-195 expressions were reduced in patients with CHD compared to that of controls (all p < 0.001). In CHD patients, lnc-UCA1 was negatively correlated with miR-26a (p < 0.001) and miR-195 (p = 0.014). Besides, lnc-UCA1 was positively correlated with Gensini score (p < 0.001), total cholesterol (p = 0.019), low-density lipoprotein cholesterol (p = 0.002), and C-reactive protein (p < 0.001), while miR-26a (p < 0.001) and miR-195 (p = 0.002) were negatively correlated with Gensini score. What's more, lnc-UCA1 was positively correlated with tumor necrosis factor (TNF)-α (p = 0.004), interleukin (IL)-1ß (p = 0.041), vascular cell adhesion molecule-1 (VCAM-1) (p = 0.010), and intercellular adhesion molecule-1 (ICAM-1) (p < 0.001). While miR-26a was negatively correlated with some of the individual inflammatory cytokines and cell adhesion molecules. CONCLUSION: Lnc-UCA1, miR-26a, and miR-195 may serve as potential biomarkers for CHD management.

Asunto(s)

Enfermedad Coronaria , MicroARNs/sangre , ARN Largo no Codificante/sangre , Anciano , Moléculas de Adhesión Celular/sangre , Enfermedad Coronaria/sangre , Enfermedad Coronaria/epidemiología , Enfermedad Coronaria/patología , Estenosis Coronaria/patología , Citocinas/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad

RESUMEN

We aimed to explore the role of microRNA 195 (miR-195) in diabetic retinopathy (DR). From January 2019 to July 2020, 50 patients with DR undergoing vitrectomy and 40 patients with idiopathic macular holes undergoing vitrectomy were selected as the observation group (OG) and control group (CG), respectively. The mRNA and protein expression levels of miR-195, SIRT1, BAX, and BCL-2 were detected in the retinal tissues obtained from the two groups during surgery. In addition, human retinal endothelial cells and human dermal microvascular endothelial cells were cultured in a high-glucose environment to detect the targeted relationship between miR-195 and SIRT1; determine the mRNA and protein expression levels of SIRT1, BAX, and BCL-2 after miR-195 knockdown; and assess the levels of cell proliferation and apoptosis. In OG, the mRNA and protein expression levels of miR-195 and BAX were high, whereas those of BCL-2 and SIRT1 were low. Moreover, we detected a targeted relationship between miR-195 and SIRT1. Conversely, miR-195 knockdown led to the downregulation of the mRNA and protein expression levels of BAX and the upregulation of the mRNA and protein expression levels of SIRT1 and BCL-2 as well as improvement in cell growth and a decrease in the apoptosis rate. miR-195 is overexpressed in DR, and its targeted relationship with SIRT1 inhibits the growth of cells in the retina and accelerates apoptosis.

Asunto(s)

Diabetes Mellitus , Retinopatía Diabética , MicroARNs , Apoptosis/genética , Retinopatía Diabética/genética , Retinopatía Diabética/metabolismo , Células Endoteliales/metabolismo , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/genética , ARN Mensajero , Sirtuina 1/genética , Sirtuina 1/metabolismo , Proteína X Asociada a bcl-2/genética

RESUMEN

Mind bomb 1 (MIB1) is a well­known E3 ubiquitin ligase. MicroRNAs (miRNAs/miRs) have been found to serve important functions in cancer cell physiology. However, the clinical significance and biological function of MIB1 and miRNAs in prostate cancer (PCa) are yet to be fully elucidated. The current study predicted the interaction between MIB1 and miR­195­5p using TargetScan, and the results were confirmed by performing a dual­luciferase reporter assay. The mRNA expression level of MIB1 and miR­195­5p in PCa and adjacent normal tissues, and PCa cell lines was detected using reverse transcription­quantitative PCR. Cell Counting Kit­8 and Transwell assays were used to measure the proliferation, and migration and invasion of VCaP and DU145 PCa cell lines, respectively, while western blot analysis was used to detect the protein expression level of MIB1. The results revealed that the mRNA expression level of MIB1 was increased, while the mRNA expression level of miR­195­5p was decreased in PCa tissues (P<0.001 and P<0.01, respectively) and in various cell lines, including PC­3 (P<0.001 and P<0.05, respectively), VCaP (P<0.001 and P<0.01, respectively), 22Rv1 (P<0.001 and P<0.05, respectively), DU145 (P<0.001 and P<0.01, respectively) and LNCaP (P<0.001 and P<0.05, respectively). miR­195­5p mimics rescued the inhibitory effects caused by knockdown of MIB1 on cell proliferation, migration and invasion in the VCaP and DU145 cell lines. In addition, MIB1 overexpression restored the miR­195­5p overexpression­induced repression of cell proliferation and invasion. The current study revealed that the MIB1 gene was an effector of cell proliferation, migration and invasion in PCa cell lines. Furthermore, miR­195­5p may regulate PCa cell proliferation and invasion by regulating MIB1, indicating its potential therapeutic application for PCa in the future.

Asunto(s)

Movimiento Celular/genética , Proliferación Celular/genética , MicroARNs/genética , Invasividad Neoplásica/genética , Neoplasias de la Próstata/genética , Ubiquitina-Proteína Ligasas/genética , Línea Celular Tumoral , Regulación Neoplásica de la Expresión Génica , Humanos , Masculino , Regulación hacia Arriba

RESUMEN

In most human primary cancers, the expression, or telomerase activity, of telomerase reverse transcriptase (TERT) is detectable. However, the mechanism ofTERTactivity within oncogenesis of thyroid cancer remains largely unknown. In this study, we identified miR-195-5p as having involvement in cell proliferation, apoptosis, and invasion in human thyroid cancer. MTT was used to measure cell proliferation, Transwell chamber was used to measure invasion. Western blotting was used to detect the expressions of TERT, PCNA, and Ki67. Target gene prediction software predicted that TERT may be the target gene of miR-195-5p. Luciferase reporting system was used to identify the targeting relationship. A significant increase of in TERT expression was observed by immunohistochemistry compared with normal tissue, however, a decrease in miR-195-5p expression using qRT-PCRand western blot compared with normal cells. Functional analysis demonstrates that miR-195-5p negatively correlated withTERTand inhibitedTERTexpression through its interaction with theTERT3'-untranslatedregion (3'-UTR). Overexpression of miR-195-5p was shown to inhibit proliferation and invasion, and promote apoptosis of CAL-62 thyroid cancer cells. miR-195-5p-mediatedeffects were rescued by the overexpression ofTERT. Altogether, our data demonstrate that miR-195-5p regulates cell proliferation, apoptosis, and invasion in human thyroid cancer viaTERT, providing evidence of a new potential therapeutic target for further investigation.

Asunto(s)

MicroARNs/genética , Telomerasa/genética , Neoplasias de la Tiroides , Apoptosis/genética , Línea Celular Tumoral , Proliferación Celular/genética , Humanos , MicroARNs/metabolismo , Invasividad Neoplásica/genética , Telomerasa/metabolismo , Neoplasias de la Tiroides/genética , Neoplasias de la Tiroides/metabolismo , Neoplasias de la Tiroides/patología

RESUMEN

Increasing evidence has indicated that microRNAs (miRNAs/miRs) play an important role in the occurrence and development of various types of cancer. The aim of the present study was to investigate the role and underlying molecular mechanisms of miR-195-5p in laryngeal cancer cell proliferation, migration and invasion. Reverse transcription-quantitative PCR (RT-qPCR) was performed to measure the expression levels of miR-195-5p in laryngeal carcinoma cell lines. The expression levels of miR-195-5p and E2F transcription factor 3 (E2F3) were modified by transfection with miR-195-5p mimics and pcDNA3.1-E2F3. A luciferase reporter assay was used to verify the association between miR-195a-5p and E2F3. Cell Counting Kit-8, cell wound healing and Transwell invasion assays were used to detect the biological functions of laryngeal cancer cells. The expression of epithelial-mesenchymal transition (EMT)-associated genes was evaluated by western blotting and RT-qPCR. The results revealed that the expression of miR-195-5p was decreased in laryngeal cancer cell lines. The overexpression of miR-195-5p inhibited the proliferation, migration, invasion and EMT of laryngeal cancer cells. Dual-luciferase reporter assays revealed that miR-195-5p could directly target E2F3 and that there was a negative association between them. E2F3 overexpression significantly attenuated the inhibitory effects of the overexpression of miR-195-5p on the proliferation, migration, invasion and EMT of laryngeal cancer cells. Collectively, the findings of the present study demonstrated that the overexpression of miR-195-5p significantly inhibited the progression of laryngeal cancer cells, and these effects may be mediated via the downregulation of the expression of E2F3.