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1.
Anticancer Res ; 40(1): 239-244, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31892572

RESUMEN

BACKGROUND: In previous studies, we demonstrated the significant role of microRNA-449a (miR-449a) in colorectal cancer with in vivo and clinical samples. The importance of miR-449a in gastric cancer is still to be elucidated. This study examined the impact of miR-449a expression in tumor tissue and serum and investigated its potential as a prognostic marker in gastric cancer. MATERIALS AND METHODS: Sixty-six patients with gastric cancer who underwent surgery were included in the study. miR-449a expression in tumor tissue and serum were investigated by real-time polymerase chain reaction analysis. The association of miR-449a expression with clinicopathological factors and patient prognosis were also investigated. RESULTS: miR-449a expression was lower in tumor tissue than non-tumor tissue. miR-449a in tumor tissue negatively correlated with the malignancy of tumor and clinical stage. Increased carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) levels were seen at significantly higher frequencies in patients with low miR-449a expression. Patients with low miR-449a expression had poorer cancer-specific survival compared to those with high miR-449a expression. The univariate analysis showed that lymphovascular invasion, increased CEA and CA19-9 and a low expression of miR-449a were associated with a poorer 5-year cancer-specific survival. miR-449a expression level in serum correlated to that in tumor tissue and was also associated with tumor malignancy. CONCLUSION: The miR-449a level in tumor tissue might be useful as a prognostic indicator for patients with gastric cancer and miR-449a in serum appears to reflect its expression in tumor tissue.


Asunto(s)
Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , Neoplasias Gástricas/genética , Anciano , Supervivencia sin Enfermedad , Femenino , Humanos , Masculino , MicroARNs/sangre , MicroARNs/metabolismo , Pronóstico , Neoplasias Gástricas/sangre
2.
Recent Results Cancer Res ; 215: 277-298, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31605235

RESUMEN

Deregulation of microRNA expression has been shown to play an important role in human malignancies. The identification of circulating-free miRNAs in biofluids a decade ago led to great enthusiasm and motivation to develop non-invasive tests based on the expression of these small non-coding RNAs. Herein, we review the progress within the field of research for identifying circulating miRNA cancer biomarkers and discuss the advantages and challenges associated with this. We also discuss the methodological and analytical variables, which may influence the final miRNA quantification and the importance of standardizing pre-analytical, analytical, and post-analytical processes in order to enable a successful translation of the results from basic research into the clinics.


Asunto(s)
Biomarcadores de Tumor/genética , MicroARNs/sangre , Neoplasias/sangre , Neoplasias/genética , Biomarcadores de Tumor/sangre , Humanos , MicroARNs/análisis , MicroARNs/genética
3.
Gene ; 724: 144156, 2020 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-31626960

RESUMEN

Cadmium is a common environmental and occupational pollutant and can produce toxic effects in a range of organs, especially in kidneys, after long-term exposure. MicroRNAs are ideal candidate biomarkers for various types of disorders, including renal diseases. In this study, we profiled the global miRNA expressions in rat kidneys using miRNA microarrays and found a collection of differentially expressed miRNAs induced by cadmium exposure. Among all of the candidate miRNAs, we identified miR-122-5p and miR-326-3p as early biomarkers for cadmium-induced nephrotoxicity. The two-miRNA signature was validated by quantitative real-time PCR in HK-2 and NRK-52E cells, rat kidney, serum and urine samples, and serum of an occupational population. Our results indicate that miR-122-5p and miR-326-3p may be potential biomarkers for cadmium exposure.


Asunto(s)
Cadmio/toxicidad , Biomarcadores Ambientales/genética , Exposición a Riesgos Ambientales/análisis , Riñón/efectos de los fármacos , MicroARNs/análisis , Adulto , Animales , Estudios de Casos y Controles , Contaminantes Ambientales/toxicidad , Femenino , Humanos , Riñón/metabolismo , Masculino , MicroARNs/sangre , Exposición Profesional/análisis , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Transcriptoma
5.
Medicine (Baltimore) ; 98(50): e17814, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31852062

RESUMEN

The purpose of our research was to evaluate diagnostic performance of serum microRNA-135a (miR-135a) in non-small cell lung cancer (NSCLC).Quantitative real time-polymerase chain reaction was employed to detect the expression serum of miR-135a in NSCLC patients and controls. The influence of serum miR-135a level on clinical characteristics of NSCLC patients was explored through the Chi-square test. Serum carcinoembryonic antigen (CEA) level was estimated via enzyme-linked immunosorbent assay. Receiver operating characteristic (ROC) curve was plotted to elucidate diagnostic roles of serum miR-135a and CEA in NSCLC.The expression level of serum miR-135a was significantly lower in NSCLC patients than in healthy controls (0.40 ±â€Š0.29 vs 1.00 ±â€Š0.40, P < .001). Moreover, miR-135a expression was related to lymph node metastasis (P = .021), tumor differentiation (P = .020), and tumor node metastasis stage (P = .031). ROC curve showed serum miR-135a level could discriminate NSCLC patients from healthy controls (P < .0001) with a corresponding cutoff value of 0.665, and a sensitivity and specificity of 81.3% and 83.1%, respectively. The area under the curve was 0.888. In diagnosis analysis on the combination of miR-135a and CEA, when its specificity was maintained at 90%, diagnosis cut-off point reached 0.678.Serum miR-135a level is significantly downregulated in NSCLC and serves as a potential diagnostic biomarker for the disease.


Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/diagnóstico , Neoplasias Pulmonares/diagnóstico , MicroARNs/sangre , Estadificación de Neoplasias/métodos , Anciano , Biomarcadores de Tumor/sangre , Carcinoma de Pulmón de Células no Pequeñas/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Neoplasias Pulmonares/sangre , Masculino , Persona de Mediana Edad , ARN Neoplásico/genética , Curva ROC , Reacción en Cadena en Tiempo Real de la Polimerasa
6.
Chem Commun (Camb) ; 55(99): 14958-14961, 2019 Dec 25.
Artículo en Inglés | MEDLINE | ID: mdl-31774422

RESUMEN

A simple method for direct detection of microRNAs (miRs) in human serum without the use of polymerase amplification is presented, achieving low miR-122 concentrations and importantly, discerning effectively single-base sequence mutations. The method is based on the capture of target miRs with synthetic peptide nucleic acid oligomers, dynamic chemical labelling, separation with quaternary amine microplatforms and detection using time-gated fluorescence imaging.


Asunto(s)
MicroARNs/sangre , Imagen Óptica/métodos , Colorantes Fluorescentes/química , Humanos , Límite de Detección
7.
Medicine (Baltimore) ; 98(44): e17710, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31689804

RESUMEN

Tropomyosin 1 (TPM1) is a protein that constitutes the sarcomere filaments and is encoded by the TPM1 gene. The aim of the present study is to investigate the correlation between the 3' untranslated region (3'UTR) single nucleotide polymorphisms (SNPs) of the TPM1 gene and dilated cardiomyopathy (DCM).A total of 245 patients with DCM and 245 healthy controls were recruited with 5 ml of venous blood. Genomic DNA was extracted to analyze the TPM1 gene rs12148828, rs11558748, rs707602, rs6738, rs7178040 loci genotypes, and the plasma miR-21 level was analyzed by reverse transcription-PCR (RT-PCR).The risk of DCM development in the rs6738 locus G allele carriers were 1.69 times more than A allele carriers (95% CI: 1.22-2.33, P = .001). Age and gender had no effect on the association of TPM1 gene SNPs with DCM risk (P > .05). The plasma miR-21 level of TPM1 gene rs6738 locus AA carriers was significantly higher than that of the AG and GG genotypes (P < .001).The SNPs of TPM1 gene rs6738 locus is associated with the risk of DCM, which may be related to the abnormal increase of miR-21 level in DCM patients, but further research is needed to prove the causal relationship between miR-21 level and DCM risk.


Asunto(s)
Regiones no Traducidas 3'/genética , Cardiomiopatía Dilatada/genética , Predisposición Genética a la Enfermedad/genética , Polimorfismo de Nucleótido Simple/genética , Tropomiosina/genética , Adulto , Anciano , Anciano de 80 o más Años , Alelos , Cardiomiopatía Dilatada/sangre , Estudios de Casos y Controles , Femenino , Genotipo , Humanos , Masculino , MicroARNs/sangre , Persona de Mediana Edad , Factores de Riesgo
8.
Cancer Sci ; 110(12): 3677-3688, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31646712

RESUMEN

5-Fluorouracil (5-FU) is a chemotherapeutic agent commonly used to treat esophageal squamous cell carcinoma (ESCC), but acquisition of chemoresistance frequently occurs and the underlying mechanisms are not fully understood. We found that microRNA (miR)-338-5p was underexpressed in ESCC cells with acquired 5-FU chemoresistance. Forced expression of miR-338-5p in these cells resulted in downregulation of Id-1, and restoration of both in vitro and in vivo sensitivity to 5-FU treatment. The effects were abolished by reexpression of Id-1. In contrast, miR-338-5p knockdown induced 5-FU resistance in chemosensitive esophageal cell lines, and knockdown of both miR-338-5p and Id-1 resensitized the cells to 5-FU. In addition, miR-338-5p had suppressive effects on migration and invasion of ESCC cells. Luciferase reporter assay confirmed a direct interaction between miR-338-5p and the 3'-UTR of Id-1. We also found that miR-338-5p was significantly downregulated in tumor tissue and serum samples of patients with ESCC. Notably, low serum miR-338-5p expression level was associated with poorer survival and poor response to 5-FU/cisplatin-based neoadjuvant chemoradiotherapy. In summary, we found that miR-338-5p can modulate 5-FU chemoresistance and inhibit invasion-related functions in ESCC by negatively regulating Id-1, and that serum miR-338-5p could be a novel noninvasive prognostic and predictive biomarker in ESCC.


Asunto(s)
Neoplasias Esofágicas/genética , Carcinoma de Células Escamosas de Esófago/genética , Proteína 1 Inhibidora de la Diferenciación/genética , MicroARNs/fisiología , Adulto , Anciano , Animales , Línea Celular Tumoral , Movimiento Celular , Resistencia a Antineoplásicos , Neoplasias Esofágicas/tratamiento farmacológico , Neoplasias Esofágicas/mortalidad , Neoplasias Esofágicas/patología , Carcinoma de Células Escamosas de Esófago/tratamiento farmacológico , Carcinoma de Células Escamosas de Esófago/mortalidad , Carcinoma de Células Escamosas de Esófago/patología , Femenino , Fluorouracilo/farmacología , Humanos , Masculino , Ratones , MicroARNs/sangre , Persona de Mediana Edad , Invasividad Neoplásica
9.
Chem Commun (Camb) ; 55(91): 13733-13736, 2019 Nov 25.
Artículo en Inglés | MEDLINE | ID: mdl-31661100

RESUMEN

The tough challenges for the convenient and quantitative determination of circulating miRNAs (cmiRNAs) in the peripheral blood are low abundance, high interference and lack of direct digital readout. Here, we developed dual-enhanced magnetobiosensors based on cascaded nucleic acid circuits, which integrate catalyzed hairpin assembly (CHA) with the hybridization chain reaction (HCR), for sensitive, portable and digital quantitative detection of circulating miRNAs in serum by a personal glucose meter (PGM).


Asunto(s)
Técnicas Biosensibles/métodos , Campos Magnéticos , MicroARNs/sangre , Ácidos Nucleicos/química , Glucosa/química , Humanos , Hibridación de Ácido Nucleico
10.
Life Sci ; 238: 116894, 2019 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-31626789

RESUMEN

AIMS: MicroRNAs (miRs) and their importance in development, normal physiology, and disease have become increasingly recognized. Our laboratory is interested in miR-29 and its effects on lung development. These studies set out to identify optimal conditions for the measurement of miR-29 in heparinized, biobanked samples and to compare isoform expression patterns. MATERIALS AND METHODS: The efficiency of three distinct heparinases were tested using reverse transcriptase polymerase chain reaction (RT-PCR): recombinant F. Heparinum heparinase I; recombinant P. heparinus heparinase II; recombinant P. heparinus heparinase III; and heparinase I (B. efferthii-derived). The effects of freeze/thaws, and the relative expression of different miR-29 isoforms were also assessed using RT-PCR. KEY FINDINGS: Our investigations determined that heparinase 1 (recombinant F. Heparinum) and 2 (recombinant P. heparinus) at 1 or 2 h incubation efficiently neutralized heparin activity and prevented interference with the PCR. Also, a single freeze/thaw did not affect the measurement of miR-29-3p but multiple freeze/thaw cycles decreased the measureable miR levels. Finally, the -3p strand was most abundantly expressed in all three isoforms in both human and mouse plasma. SIGNIFICANCE: Our findings illustrate that specific conditions need to be optimized for the particular miR and the type of sample being tested.


Asunto(s)
Bancos de Muestras Biológicas/normas , Heparina/sangre , MicroARNs/sangre , Animales , Estudios de Cohortes , Flavobacterium/enzimología , Liasa de Heparina/metabolismo , Humanos , Lactante , Ratones , Ratones Endogámicos C57BL , Reacción en Cadena de la Polimerasa , Polisacaridoliasas/metabolismo , Proteínas Recombinantes/metabolismo
11.
Cell Prolif ; 52(6): e12704, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31621141

RESUMEN

Intervertebral disc degeneration (IDD) is a common cause of low back pain, which inflicts more global disability than any other condition. Although IDD was deemed to be a natural process that comes with ageing, a growing body of evidence suggested that both genetic and environmental factors could modify the development of IDD. In this connection, aberrant function of nucleus pulposus cells has been implicated in IDD pathogenesis. Circular RNAs are a novel class of endogenous non-coding RNAs that play crucial regulatory roles in diverse cellular processes. Recently, deregulation of circRNAs in nucleus pulposus cells was found to functionally participate in IDD development. In this review, we summarize the current knowledge regarding the deregulation of circRNAs in IDD in relation to their actions on nucleus pulposus cell functions, including cell proliferation, apoptosis and extracellular matrix synthesis/degradation. The potential clinical utilities of circRNAs as therapeutic targets for the management of IDD are also discussed.


Asunto(s)
Degeneración del Disco Intervertebral/sangre , Disco Intervertebral/patología , MicroARNs/sangre , Núcleo Pulposo/metabolismo , Proliferación Celular/genética , Humanos , Transducción de Señal/genética
12.
Virol J ; 16(1): 116, 2019 10 07.
Artículo en Inglés | MEDLINE | ID: mdl-31590680

RESUMEN

BACKGROUND: Finding new biomarkers for the early detection of cervical cancer is an essential requirement in this field. In this study, we aimed to evaluate the expression level of potential biomarkers in progression of cervical cancer in patients with cervical cancer compared to normal subjects. METHODS: The expression levels of tissue and serum miRNAs, including miR-9, miR-192 and miR-205, were investigated in 36 normal, 18 precancer, and 18 cervical cancer samples using real-time polymerase chain reaction. RESULTS: The results showed the higher significant expressions of miR-9, miR-192 and miR-205 in the tissue of cancer samples than those in the normal samples. Moreover, the miR-192 and miR-205 expression were significantly increased in the cancer group in comparison with the precancer group. Examination of serum samples revealed the increase in the expression level in the cancer groups than in the normal samples, for miR-9, miR-192 and miR-205 and the expressions of miR-9, miR-192 and miR-205 were significantly up-regulated in the precancer group in comparison with the normal group. Also the expression of miR-205 was remarkably increased in the cancer group in comparison with the precancer group. The receiver operating characteristic (ROC) analyses showed the highest area under the curve value for miR-192. CONCLUSIONS: Given the increased expression level of miR-192 in cancer and in precancerous tissue and serum compared with the normal tissue and serum validated by analysing the ROC curve, miR-192 can be used as potential biomarker for the early detection of cervical cancer.


Asunto(s)
Biomarcadores de Tumor/sangre , Detección Precóz del Cáncer , MicroARNs/sangre , MicroARNs/genética , Neoplasias del Cuello Uterino/diagnóstico , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Estudios de Casos y Controles , Cuello del Útero/patología , Cuello del Útero/virología , Femenino , Humanos , Persona de Mediana Edad , Curva ROC , Regulación hacia Arriba , Adulto Joven
13.
Medicine (Baltimore) ; 98(39): e17297, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31574853

RESUMEN

As a modifiable risk factor for cardiovascular disease, presence of hypertension (HT) necessitates the awareness of asymptomatic organ damage (AOD). The aim of this study was to measure plasma micro RNA-21 (miR-21) and the parameters that reflect AOD such as carotid intima-media thickness (CIMT), microalbuminuria (MAU) in hypertensive patients compared with healthy controls. In addition, the aim of this study was to evaluate plasma miR-21 levels in HT patients with AOD.This study was designed as a cross-sectional observational study. The study includes 2 groups: 32 patients with HT and 32 healthy controls. First, we compared these 2 groups. Then, to underline the relationship between plasma miR-21 and HT, hypertensive patients were divided into 2 groups: with AOD and without AOD.Sixteen patients with HT had AOD. MiR-21 levels significantly correlated with clinical systolic and diastolic blood pressure, MAU, C-reactive protein, and CIMT. CIMT, miR-21, and MAU levels were significantly higher in patients with AOD.Our study showed increased miR-21 levels in HT patients with AOD.


Asunto(s)
Albuminuria , Enfermedades Cardiovasculares , Hipertensión , MicroARNs/sangre , Adulto , Albuminuria/diagnóstico , Albuminuria/etiología , Enfermedades Asintomáticas/epidemiología , Presión Sanguínea/fisiología , Proteína C-Reactiva/análisis , Enfermedades Cardiovasculares/diagnóstico , Enfermedades Cardiovasculares/epidemiología , Enfermedades Cardiovasculares/etiología , Grosor Intima-Media Carotídeo , MicroARN Circulante/análisis , Correlación de Datos , Estudios Transversales , Femenino , Humanos , Hipertensión/complicaciones , Hipertensión/diagnóstico , Hipertensión/genética , Hipertensión/fisiopatología , Masculino , Persona de Mediana Edad , Turquia/epidemiología
14.
Medicine (Baltimore) ; 98(42): e17335, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31626089

RESUMEN

BACKGROUND: Previous studies have shown that microRNA-32 (miRNA-32) is an exosome microRNA that affects the proliferation and metastasis of non-small cell lung cancer (NSCLC) cells. In this study, our goal was to assess the expression of plasma microRNA-32 and its potential as a biomarker to predict the tumor response and survival of patients with NSCLC undergoing platinum-based chemotherapy. METHODS: Plasma microRNA-32 levels before and after 1 cycle of platinum-based chemotherapy in 43 patients with NSCLC were measured using a quantitative real-time polymerase chain reaction assay (qPCR). In addition, the demographic and survival data of the patients were collected for analysis. RESULTS: A significant correlation was observed between the changes in microRNA-32 levels before and after 1 chemotherapy cycle and the treatment response (P = .035). In addition, Kaplan-Meier analysis showed that the level of microRNA-32 after 1 chemotherapy cycle was significantly correlated with the prognosis of patients. The median progression-free survival (P = .025) and overall survival (P = .015) of patients with high microRNA-32 levels (≥7.73) after 1 chemotherapy cycle was 9 and 21 months, respectively. In contrast, the median survival of patients with low microRNA-32 levels (<7.73) was 5 and 10 months, respectively. CONCLUSIONS: The plasma levels of microRNA-32 correlated with the efficacy of platinum-based chemotherapy and survival, indicating that microRNA-32 may be useful for predicting the effectiveness of platinum-based chemotherapy and prognosis in NSCLC.


Asunto(s)
Antineoplásicos/uso terapéutico , Cisplatino/uso terapéutico , Neoplasias Pulmonares/genética , MicroARNs/genética , Carcinoma Pulmonar de Células Pequeñas/genética , Anciano , Biomarcadores de Tumor/genética , Estudios Controlados Antes y Después , Femenino , Humanos , Estimación de Kaplan-Meier , Neoplasias Pulmonares/sangre , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/mortalidad , Masculino , MicroARNs/sangre , Persona de Mediana Edad , Terapia Neoadyuvante , Modelos de Riesgos Proporcionales , Reacción en Cadena en Tiempo Real de la Polimerasa , Carcinoma Pulmonar de Células Pequeñas/sangre , Carcinoma Pulmonar de Células Pequeñas/tratamiento farmacológico , Carcinoma Pulmonar de Células Pequeñas/mortalidad , Resultado del Tratamiento
15.
Eur J Endocrinol ; 181(5): 525-537, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31536965

RESUMEN

Objective: To evaluate the effect of insulin resistance in obesity on the expression in whole blood of mRNA and miRNA affecting bone homeostasis as well as to estimate the influence of oral glucose load (OGTT) on serum osteocalcin concentration in obese individuals with and without insulin resistance. Design: Cross-sectional study. Methods: Carboxylated (cOC), undercarboxylated (ucOC) and total osteocalcin were measured by ELISA in the serum of obese subjects with insulin resistance (n = 41) and obese subjects without insulin resistance (n = 41) (control group) during OGTT. Analysis of gene expression (microarray) and miRNAs (real-time PCR) was performed in venous blood (representating samples) collected before OGTT from obese with insulin resistance and controls. Results: Obese subjects with insulin resistance (higher HOMA-IR and lower oral glucose insulin sensitivity index) presented significantly increased expression of WNT signalling inhibitors (DKK1, DKK2, SOST, SFRP1) and downregulation of the key factor in WNT signalling - ß catenin participating in osteoblasts differentiation. Expression of miRNA involved in osteoblastogenesis was also inhibited (miR-29b, miR-181a, miR-210, miR-324-3p). During OGTT, contrary to the control group, subjects with insulin resistance presented suppression of cOC and total OC decrease after 1 and 2 h of oral glucose load. Conclusions: Obese subjects with insulin resistance may have defects in osteoblastogenesis that was demonstrated via key signalling molecules mRNA downregulation, and increased expression of WNT antagonists as well as inhibition of expression of miRNA participating in the regulation of osteoblast differentiation. Disturbed osteoblastogenesis in insulin-resistant subjects results in the suppression of blood carboxylated and total osteocalcin decrease during OGTT.


Asunto(s)
Remodelación Ósea/fisiología , Resistencia a la Insulina/fisiología , MicroARNs/sangre , ARN Mensajero/sangre , Adulto , Estudios Transversales , Ensayo de Inmunoadsorción Enzimática , Femenino , Prueba de Tolerancia a la Glucosa , Humanos , Masculino , Análisis por Micromatrices , Persona de Mediana Edad , Obesidad/etiología , Obesidad/metabolismo , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Vía de Señalización Wnt/fisiología
16.
Eur J Endocrinol ; 181(5): 565-577, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31539877

RESUMEN

Design: Gestational diabetes mellitus (GDM) is one of the most common pregnancy complications and its prevalence is constantly rising worldwide. Diagnosis is commonly in the late second or early third trimester of pregnancy, though the development of GDM starts early; hence, first-trimester diagnosis is feasible. Objective: Our objective was to identify microRNAs that best distinguish GDM samples from those of healthy pregnant women and to evaluate the predictive value of microRNAs for GDM detection in the first trimester. Methods: We investigated the abundance of circulating microRNAs in the plasma of pregnant women in their first trimester. Two populations were included in the study to enable population-specific as well as cross-population inspection of expression profiles. Each microRNA was tested for differential expression in GDM vs control samples, and their efficiency for GDM detection was evaluated using machine-learning models. Results: Two upregulated microRNAs (miR-223 and miR-23a) were identified in GDM vs the control set, and validated on a new cohort of women. Using both microRNAs in a logistic-regression model, we achieved an AUC value of 0.91. We further demonstrated the overall predictive value of microRNAs using several types of multivariable machine-learning models that included the entire set of expressed microRNAs. All models achieved high accuracy when applied on the dataset (mean AUC = 0.77). The significance of the classification results was established via permutation tests. Conclusions: Our findings suggest that circulating microRNAs are potential biomarkers for GDM in the first trimester. This warrants further examination and lays the foundation for producing a novel early non-invasive diagnostic tool for GDM.


Asunto(s)
MicroARN Circulante/sangre , Diabetes Gestacional/sangre , Diabetes Gestacional/diagnóstico , Tejido Adiposo/química , Adulto , Estudios de Casos y Controles , Diagnóstico Precoz , Femenino , Humanos , Aprendizaje Automático , MicroARNs/sangre , Placenta/química , Valor Predictivo de las Pruebas , Embarazo , Primer Trimestre del Embarazo , Reproducibilidad de los Resultados
17.
Analyst ; 144(19): 5856-5865, 2019 Sep 23.
Artículo en Inglés | MEDLINE | ID: mdl-31482867

RESUMEN

Exosome-containing microRNAs (exomiRs) can be employed as potential biomarkers for tumor diagnosis and have drawn much attention in the past few years. However, the separation of exosomes and the detection of exomiRs are still inconvenient or even difficult to implement. Thus, it is important to develop a simple, accurate, and reliable strategy for the separation of exosomes and the biomedical analysis of exomiRs. Herein, a novel exosome-specific tumor diagnosis strategy was constructed by integrating the rapid magnetic exosome-enrichment platform and the Ru(bpy)32+-polymer amplified electrochemiluminescence (ECL) strategy. This strategy realized the rapid and efficient capture of tumor-derived exosomes through a biological-affinity identification platform of the EpCAM antibody. The biomedical analysis of exomiRs achieved a preferable specificity and high sensitivity of 103 particles. Furthermore, we investigated the performance index for clinical blood samples from tumor patients; the results indicated that the exosome-specific tumor diagnosis strategy readily and consistently responded to exomiRs. These results indicated that the exosome-specific tumor diagnosis strategy provided new opportunities for the sensitive and efficient analysis of tumor-derived exomiRs. This strategy greatly simplified the biomedical analysis process and established the non-destructive detection mode of fluid biopsy for tumors.


Asunto(s)
Biomarcadores de Tumor/sangre , Exosomas/metabolismo , MicroARNs/sangre , Neoplasias/diagnóstico , Células A549 , Biomarcadores de Tumor/química , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , Células Hep G2 , Humanos , Luminiscencia , Células MCF-7 , Nanopartículas de Magnetita/química , MicroARNs/química , Neoplasias/ultraestructura , Compuestos Organometálicos/química , Polímeros/química , Sensibilidad y Especificidad
18.
Mikrochim Acta ; 186(10): 669, 2019 09 05.
Artículo en Inglés | MEDLINE | ID: mdl-31489499

RESUMEN

A highly sensitive fluorometric method is described for the determination of let-7a microRNA. It is based on the use of target-triggered cascade signal amplification that involves the use of (a) catalytic hairpin assembly (CHA), (b) exonuclease-assisted signal amplification (EASA), and (c) DNA-templated fluorescent silver nanoclusters (DNA-AgNCs) having excitation/emission maxima at 535/616 nm. The CHA reaction is initiated by the hybridization of the microRNA with hairpin probe HP1. The opening of HP1 results in the assembly of HP1 and another hairpin probe (HP2) to form a duplex. This releases the microRNA that which initiates another CHA reaction. The HP1-HP2 duplex binds to hairpin probe HP3 to form a stable Y-shaped junction structure HP2-HP1-HP3. The Y-shaped junction structure is cleaved by λ exonuclease to recycle the HP1-HP2 duplex to initiate the EASA reaction. This generates a large number of single-stranded reporter sequences. These act as scaffolds for the synthesis of fluorescent AgNCs by reduction of Ag (I) ions. A remarkably amplified fluorescent signal is observed whose intensity increases linearly in the 1 fM to 10 nM let-7a microRNA concentration range. The detection limit is 0.89 fM. The method can well discriminate let-7a microRNA from other microRNAs of the same family. Graphical Abstract Schematic representation of a fluorometric method based on target-triggered cascade signal amplification and DNA-templated silver nanoclusters (DNA-AgNCs) for sensitive detection of microRNA (miRNA).


Asunto(s)
ADN/química , Nanopartículas del Metal/química , MicroARNs/sangre , Sondas Moleculares/química , Técnicas de Amplificación de Ácido Nucleico , Plata/química , Técnicas Biosensibles , Humanos , MicroARNs/análisis , Espectrometría de Fluorescencia
19.
Anal Chim Acta ; 1082: 176-185, 2019 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-31472706

RESUMEN

We report herein a novel multiple electrochemical aptasensor based on covalent-organic framework (COF) for sensitive and simultaneous detection of miRNA 155 and miRNA 122, by using shell-encoded gold nanoparticles (Au NPs) as signal labels (AgNCs@AuNPs and Cu2O@AuNPs, respectively, NCs = nanoclusters). A new COF nanowire was synthesized via condensation polymerization of 1,3,6,8-tetra(4-carboxylphenyl)pyrene and melamine (represented by TBAPy-MA-COF-COOH) for multiple aptasensor fabrication. The nanowire was then used as a platform for anchoring single-strand DNA (ssDNA), which was hybridized with the complementary aptamer (cApt) probes of miRNA 155 and miRNA 122. AgNCs@AuNPs and Cu2O@AuNPs modified with cApts show separated differential pulse voltammetry (DPV) peaks at 0.08 and -0.1 V, respectively. The signal labels immobilized with cApts were released from the hybridized DNA complex and bound to their corresponding targets when contacting miRNAs. This phenomenon results in the substantial decline of the DPV peak current density of the signal labels. The developed TBAPy-MA-COF-COOH-based aptasensor has superior performance for sensing miRNA 155 and miRNA 122 simultaneously, with ultrasensitive low detection limits of 6.7 and 1.5 fM (S/N = 3), respectively, a wide linear range of 0.01-1000 pM, and high selectivity and applicability for serum samples. The proposed TBAPy-MA-based aptasensor demonstrates potential for simultaneous detection of multiple cancer biomarkers by replacing other ssDNA and aptamer strands.


Asunto(s)
Nanopartículas del Metal/química , MicroARNs/sangre , Nanocables/química , Polímeros/química , Aptámeros de Nucleótidos/química , Aptámeros de Nucleótidos/genética , Técnicas Biosensibles/métodos , ADN de Cadena Simple/química , ADN de Cadena Simple/genética , Oro/química , Humanos , Límite de Detección , Hibridación de Ácido Nucleico , Pirenos/química , Reproducibilidad de los Resultados , Triazinas/química
20.
Vet Res Commun ; 43(4): 231-238, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31473888

RESUMEN

This study was performed to evaluate the hepatocyte-derived microRNA (miR)-122 as novel diagnostic biomarker in canine lymphoma. Fifteen dogs were enrolled in this study. Dogs presented at Small Animal Teaching Hospital, Faculty of Veterinary Medicine, Cairo University. Dogs were divided into 8 clinically healthy dogs act as control and 7 clinically ill dogs. All dogs were subjected to clinical, ultrasonographic, hemato-biochemical and ultrasound-guided fine-needle biopsy for cytological and histopathological investigations. On the basis of these results, 7 dogs were found to be suffering from multicentric lymphoma involving liver. Serum hepatocyte-derived miRA-122 was determined by real-time quantitative polymerase chain reaction in all dogs. Multicentric lymphoma involving liver manifested by inappetance for several days, depression and peripheral lymphadenopathy. Hematological examination showed significant lymphocytosis. Serum biochemical analysis revealed significant increase in ALT, AST, ALP compared to control dogs. Ultrasonography revealed hypoechoic lymphoid aggregation at area of "porta hepatis" and circumscribed hypoechoic nodule interrupt liver parenchyma. Cytology revealed infiltration of liver tissue by lymphoblast cells and histopathology revealed diffuse infiltration of hepatic sinusoids and portal area by uniform population of small lymphocytes. Serum miRNA-122 analysis showed a significant increase represented as 9.00 fold in canine multicentric lymphoma involving liver. Serum hepatocyte-derived miRNA-122 is of diagnostic value, non invasive, stable and easily measurable blood biomarker for the detection of hepatocellular injury in dogs with multicentric lymphoma involving liver.


Asunto(s)
Enfermedades de los Perros/sangre , Enfermedades de los Perros/fisiopatología , Regulación Neoplásica de la Expresión Génica , Hepatopatías/veterinaria , Hígado/patología , Linfoma/veterinaria , MicroARNs/sangre , Animales , Enfermedades de los Perros/diagnóstico , Perros , Hepatocitos/patología , Hepatopatías/sangre , Hepatopatías/complicaciones , Hepatopatías/diagnóstico , Linfoma/sangre , Linfoma/complicaciones , Linfoma/diagnóstico , MicroARNs/genética
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