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1.
Clinics (Sao Paulo) ; 78: 100155, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36681070

RESUMEN

FOXO3a dysregulation is frequently implicated in tumorigenesis, and its inhibition can occur by several molecular mechanisms. Among these, post-transcriptional suppression by miRNAs has been associated with various cancers initiation. Here, we assessed the expression profiles of the most relevant miRNAs for breast tumorigenesis, using Luminal A (LA) and Triple-Negative (TN) breast cancer from Brazilian patients, by the quantitative real time-PCR method. Their potential prognostic role for the patients was also evaluated. We identified the miRNAs miR-96-5p and miR-182-5p, de-scribed as negative regulators of FOXO3A, with differential expression both in LA and TN tumors when compared to normal tissue. The miR-96-5p and miR-182-5p miRNAs were upregulated in LA (7.82 times, p < 0.005; 6.12 times, p < 0.005, respectively) and TN breast cancer samples (9.42 times, p < 0.0001; 8.51 times, p < 0.0001) compared to normal tissues. The samples with higher miR-96-5p and miR-182-5p expression (FR ≥ 4) were submitted for FOXO3a immunostaining. Reduced protein detection was observed in all of the tumors compared to normal tissues. The most prominent miRNA expression and FOXO3a protein suppression were observed in TN samples (p < 0.001), indicating the relevant role of these molecules in this tumor biology and clinical behavior. Our results corroborate the literature regarding to the relevance of FOXO3a in the breast cancer, and they open new perspectives for alternative target therapy options for Brazilian patients expressing both FOXO3a and its regulatory miRNAs.


Asunto(s)
Neoplasias de la Mama , MicroARNs , Neoplasias de la Mama Triple Negativas , Humanos , Femenino , Neoplasias de la Mama Triple Negativas/genética , Neoplasias de la Mama Triple Negativas/patología , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Brasil , Biomarcadores de Tumor/genética , MicroARNs/genética , MicroARNs/metabolismo , Pronóstico , Carcinogénesis , Regulación Neoplásica de la Expresión Génica , Perfilación de la Expresión Génica
2.
Ren Fail ; 45(1): 2121929, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36695327

RESUMEN

BACKGROUND: Diabetic kidney disease (DKD) is one of the most common chronic complications of type 2 diabetes mellitus (T2DM), and it is particularly important to identify a high-quality method for evaluating disease progression. Urinary exosomes contain microRNA that might promise early diagnostic and monitoring markers of DKD. The present study aimed to identify novel exosome-related markers associated with inflammation and fibrosis to assess the progression of DKD. METHOD: Exosomes were extracted from the urine of 83 participants to determine the expression levels of miRNA-615-3p and miRNA-3147 in 20 healthy people, 21 patients with T2DM and 42 patients with DKD, as determined by RT-qPCR. The circulating expression level of TGF-ß1 was detected by ELISA. Serum Cystatin C was measured by a latex-enhanced immunoturbidimetric method. The correlation analyses were performed for all clinical and laboratory parameters. RESULT: The expression level of urinary exosomal miRNA-615-3p in DKD patients was significantly higher than that in the control group and the T2DM group by RT-qPCR. The expression of miRNA-3147 showed an upward trend in the three groups of subjects, but it was not statistically significant. The urinary exosomal miRNA-615-3p was positively correlated with serum Cystatin C, plasma TGF-ß1, creatinine, BUN, PCR and 24-h urine protein, and negatively correlated with eGFR and albumin. The diagnostic efficacy of urinary exosomal miRNA-615-3p combined with the ACR was higher than that of ACR alone. CONCLUSIONS: Urinary exosomal miRNA-615-3p may be used as a novel biomarker for evaluating the progression of DKD, and may be involved in the process of inflammation and fibrosis in DKD. The combined diagnosis of urinary exosomal miRNA-615-3p and ACR may be used as more stable and sensitive diagnostic criteria for DKD.


Asunto(s)
Diabetes Mellitus Tipo 2 , Nefropatías Diabéticas , MicroARNs , Humanos , MicroARNs/orina , Cistatina C , Factor de Crecimiento Transformador beta1 , Biomarcadores , Inflamación , Fibrosis
3.
Sci Rep ; 13(1): 1440, 2023 Jan 25.
Artículo en Inglés | MEDLINE | ID: mdl-36697494

RESUMEN

MicroRNAs are small molecules that play a crucial role in regulating a woman's reproductive system. The present study evaluates the expression of miR-21 in the serum, follicular fluid (FF), and cumulus cells (CCs) and their association with oocyte maturity and embryo quality in women undergoing intracytoplasmic sperm injection. Women subjects were divided into the case (54 Patients with female factor infertility) and control groups (33 patients with male factor infertility). The level of miR-21 was measured using Real-Time PCR. The level of miR-21 was significantly lower in the CCs, FF, and serum in the case compared to the control group (p < 0.05). MiR-21 abundance was higher in FF and CCs samples than in serum. Furthermore, there was a significant increase in CCs to FF in the case group (p < 0.05). A significant decrease in oocyte count, MII oocytes, and percentage of mature oocytes were observed in the case group (p < 0.05). The expression of miR-21 in FF and CCs was positively related to oocyte maturation, but no correlation with embryo development was observed. This study found that miR-21 is expressed less in women with female factor infertility, and human oocytes' development is crucially affected by the expression of miR-21. Therefore, miR-21 could provide new helpful biomarkers of oocyte maturity.


Asunto(s)
Infertilidad Femenina , MicroARNs , Humanos , Masculino , Femenino , Inyecciones de Esperma Intracitoplasmáticas , Semen/metabolismo , Oocitos/metabolismo , Líquido Folicular/metabolismo , Infertilidad Femenina/diagnóstico , Infertilidad Femenina/genética , Infertilidad Femenina/terapia , MicroARNs/genética , MicroARNs/metabolismo
4.
Shock ; 59(1): 91-98, 2023 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-36609501

RESUMEN

ABSTRACT: Ischemic postconditioning (I/Post) reduces I/R injury by activating endogenous cardioprotection mechanisms, such as the JAK/signal transducer and activator of transcription 3 (STAT3) and PI3K/Akt pathways, which offer a traditional approach to myocardial protection. According to a previous study, cardioprotection by I/Post may be lost in aged mice, and in our previous research, hypoxic postconditioning (H/Post) lacked a protective effect in senescent cardiomyocytes, which was associated with low expression of long noncoding RNA H19. The N6-methyladenosine (m 6 A) modification is a dynamic and reversible process that has been confirmed to play a role in cardiovascular diseases. However, the mechanisms of m 6 A modification in myocardial I/Post remain to be explored. Neonatal cardiomyocytes were isolated from 2-day-old Sprague-Dawley rats, and senescence was induced by d -galactose, followed by stimulation of hypoxia-reoxygenation and H/Post. Hypoxic injury was evaluated by cell viability and the Bcl-2/Bax protein ratio. Total m 6 A levels were measured using a colorimetric m 6 A RNA Methylation Quantification Kit, and the m 6 A modified and differentially expressed mRNA was determined by MeRIP (methylated RNA immunoprecipitation). We found that H/Post increased m 6 A methylation and decreased RNA mA demethylase alkB homolog 5 (ALKBH5) expression in aged cardiomyocytes. Furthermore, ALKBH5 knockdown exacerbated injury following H/Post in senescent cardiomyocytes. In addition, ALKBH5 regulated STAT3 expression by mediating its m 6 A modification and long noncoding RNA H19/miR-124-3p. ALKBH5 also alleviated the H/Post injury induced by the low expression of STAT3 in senescent cardiomyocytes.


Asunto(s)
MicroARNs , ARN Largo no Codificante , Ratas , Ratones , Animales , Miocitos Cardíacos/metabolismo , Ratas Sprague-Dawley , Galactosa/farmacología , Fosfatidilinositol 3-Quinasas/metabolismo , Factor de Transcripción STAT3/genética , Factor de Transcripción STAT3/metabolismo , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Hipoxia/metabolismo , MicroARNs/metabolismo
5.
ACS Sens ; 8(1): 317-325, 2023 Jan 27.
Artículo en Inglés | MEDLINE | ID: mdl-36617728

RESUMEN

This work presents a photocurrent-polarity-switching-based photoelectrochemical (PEC) biosensing platform for ultrasensitive detection of microRNA-21 (miR-21) through target-triggered catalytic hairpin assembly (CHA) for modulation of methylene blue (MB) and ferrocene (Fc) positional configurations using double-shelled Cu-doped ZnS nanocages (NCs)-Au nanoparticles (NPs) as photoactive materials. In the presence of miR-21, the assembly of MB-labeled HP1 and Fc-labeled HP2 leads to the generation of a large amount of double-stranded DNA (HP1-HP2), which pushes MB away from the electrode surface and brings Fc close to the electrode surface, resulting in effectively quenching the enhanced PEC signal to activate the photocurrent-polarity-switching system. Benefiting from the distance-controllable strategy, the designed PEC bioanalysis can effectively eliminate false-positive and false-negative signals due to the change of different signal expression patterns (from traditional the "signal-on" mode to the photocurrent-polarity-switching mode), thereby significantly improving the sensing specificity and sensitivity. The proposed PEC sensing system exhibited satisfying photocurrent responses toward target miR-21 within the working range from 1.0 fM to 1 nM at a low limit of detection (LOD) of 0.58 fM. More importantly, we demonstrated the successful integration of the proposed PEC biosensor with a handheld wireless device for instant detection of miR-21 concentrations in practical samples.


Asunto(s)
Técnicas Biosensibles , Nanopartículas del Metal , MicroARNs , Oro , Técnicas Biosensibles/métodos , Azul de Metileno , Técnicas Electroquímicas/métodos , MicroARNs/análisis , Proteínas Cromosómicas no Histona
6.
Anticancer Res ; 43(2): 695-706, 2023 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-36697090

RESUMEN

BACKGROUND/AIM: Non-invasive circulating tumor biomarkers in liquid biopsy, such as microRNAs (miRNA), provide for better personalization of treatment strategies. The aim of our study was to assess the prognosis of patients with melanoma undergoing tumor resection with curative intent based on analysis of selected circulating miRNAs. PATIENTS AND METHODS: A total of 22 patients with stage I to III melanoma were enrolled into this prospective study. Plasma samples were obtained pre-surgery and early post-surgery from peripheral blood draws. A panel of 23 candidate miRNAs was designed and expression of miRNAs were analyzed by reverse transcription-quantitative polymerase chain reaction with exogenous reference control cel-miR-39-3p. RESULTS: Higher preoperative expression levels of miR-99a (p=0.008), miR-320 (p=0.009), miR-1908 (p=0.001), miR-494 (p=0.018) and miR-4487 (p=0.048) were associated with a shorter disease-free interval. Similarly, higher preoperative plasma levels of miR-99a (p=0.017), miR-221 (p=0.026), miR-320 (p=0.016), miR-494 (p=0.009), miR-1260 (p=0.026) and miR-1908 (p=0.024) were associated with worse overall survival. No significant differences between pre- and postoperative plasma miRNA levels were observed. CONCLUSION: Liquid biopsy is a minimally-invasive approach which can lead to a better understanding of cancer behavior and offers the possibility of precise patient prognosis, allowing selection of the most appropriate treatment. Our study showed that preoperative plasma levels of miR-99a, miR-221, miR-320, miR-494, miR-1908 and miR-4487 were associated with disease-free interval and overall survival of patients with early-stage melanoma. This approach may help in decision-making about the appropriateness of modern adjuvant treatment administration in patients with resectable melanoma.


Asunto(s)
MicroARN Circulante , Melanoma , MicroARNs , Humanos , MicroARNs/metabolismo , Estudios Prospectivos , Pronóstico , Biomarcadores de Tumor/genética , Melanoma/genética , Melanoma/cirugía , Perfilación de la Expresión Génica
7.
Am J Pathol ; 193(2): 233-245, 2023 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-36697118

RESUMEN

As a common type of head and neck squamous cell carcinoma, oral squamous cell carcinoma (OSCC) is a lethal and deforming disease. Long noncoding RNAs have emerged as critical modulators in different malignancies. However, the role of fucosyltransferase 8 antisense RNA 1 (FUT8-AS1) in OSCC still remains elusive. In this study, quantitative RT-PCR and Western blot were used for the measurement of RNAs and proteins. Mechanism assays explored the putative correlation among genes. In vitro assays evaluated the changes in OSCC cell malignant phenotype, whereas in vivo assays highlighted the influence of FUT8-AS1 on tumor growth. FUT8-AS1, aberrantly up-regulated in OSCC tissues and cells, could exacerbate OSCC cell malignant behaviors. The cancerogenic property of FUT8-AS1 in OSCC was further confirmed via animal experiments. Furthermore, FUT8-AS1 enhanced the expression of transcription factor 4 (TCF4) via sponging miR-944 and recruiting fused in sarcoma (FUS), thus affecting OSCC cell biological behaviors via modulation on Wnt/ß-catenin signaling activity. In addition, TCF4 was validated as the transcriptional activator of FUT8-AS1. To conclude, TCF4-mediated FUT8-AS1 could exacerbate OSCC cell malignant behaviors and facilitate tumor growth via modulation on miR-944/FUS/TCF4.


Asunto(s)
Carcinoma de Células Escamosas , Neoplasias de Cabeza y Cuello , MicroARNs , Neoplasias de la Boca , ARN Largo no Codificante , Sarcoma , Animales , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Carcinoma de Células Escamosas/patología , ARN sin Sentido , Vía de Señalización Wnt/genética , Retroalimentación , Factor de Transcripción 4/genética , Factor de Transcripción 4/metabolismo , Neoplasias de la Boca/patología , Progresión de la Enfermedad , Línea Celular Tumoral , MicroARNs/genética , MicroARNs/metabolismo , Sarcoma/genética , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Proliferación Celular/genética , Regulación Neoplásica de la Expresión Génica
8.
Anal Chim Acta ; 1243: 340811, 2023 Feb 22.
Artículo en Inglés | MEDLINE | ID: mdl-36697176

RESUMEN

To establish protein enzyme-free and simple approach for sensitive detection of single nucleotide polymorphisms (SNPs), the nucleic acid amplification reactions were developed to reduce the dependence on protein enzymes (polymerase, endonuclease, ligase). These methods, while enabling highly amplified analysis for the short sequences, cannot be generalized to long genomic sequences. Herein, we develop a protein enzyme-free and general SNPs assay based on asymmetric MNAzyme probes. The multi-arm probe (MNAzyme-9M-13) with two asymmetric recognition arms, containing a short (9 nt) and a long (13 nt) arm, is designed to detect EGFR T790 M mutation (MT). Owing to the excellent selectivity of short recognition arm, MNAzyme-9M-13 probe can efficiently avoid interferences from wild-type target (WT) and various single-base mutations. Through a one-pot mixing, MNAzyme-9M-13 probe enables the sensitive detection of MT, without protein enzyme or multi-step operation. The calculated detection limit for MT is 0.59 nM and 0.83%. Moreover, this asymmetric MNAzyme strategy can be applied for SNPs detection in long genomic sequences as well as short microRNAs (miRNAs) only by changing the low-cost unlabeled recognition arms. Therefore, along with simple operation, low-cost, protein enzyme-free and strong versatility, our asymmetric MNAzyme strategy provides a novel solution for SNPs detection and genes analysis.


Asunto(s)
Técnicas Biosensibles , MicroARNs , Polimorfismo de Nucleótido Simple , Técnicas Biosensibles/métodos , Límite de Detección
9.
Mol Cells ; 46(1): 21-32, 2023 Jan 31.
Artículo en Inglés | MEDLINE | ID: mdl-36697234

RESUMEN

MicroRNAs (miRNAs) play cardinal roles in regulating biological pathways and processes, resulting in significant physiological effects. To understand the complex regulatory network of miRNAs, previous studies have utilized massivescale datasets of miRNA targeting and attempted to computationally predict the functional targets of miRNAs. Many miRNA target prediction tools have been developed and are widely used by scientists from various fields of biology and medicine. Most of these tools consider seed pairing between miRNAs and their mRNA targets and additionally consider other determinants to improve prediction accuracy. However, these tools exhibit limited prediction accuracy and high false positive rates. The utilization of additional determinants, such as RNA modifications and RNA-binding protein binding sites, may further improve miRNA target prediction. In this review, we discuss the determinants of functional miRNA targeting that are currently used in miRNA target prediction and the potentially predictive but unappreciated determinants that may improve prediction accuracy.


Asunto(s)
MicroARNs , MicroARNs/genética , MicroARNs/metabolismo , ARN Mensajero/genética , Biología Computacional/métodos
10.
Artículo en Inglés | MEDLINE | ID: mdl-36674009

RESUMEN

(1) Background: The declined function of peripheral circulating endothelial progenitor cells (EPCs) in aging individuals resulted in decreased endothelial cell regeneration and vascular endothelial function. Improving EPCs function in aging individuals plays an important role in preventing cardiovascular diseases. (2) Methods: Thirty aged (18-month-old) male Sprague-Dawley rats were randomly divided into control and exercise groups. An aerobic exercise intervention was performed 5 days/week for 8 weeks. EPCs functions, miR-21-5p, and TSP-1 expressions were detected after the intervention. The senescence rate, proliferation, and migration of EPCs were examined after overexpression of miR-21-5p and inhibition of TSP-1 expression. (3) Results: The senescence rate, proliferation, and migration of EPCs in exercise groups were significantly improved after exercise intervention. The miR-21-5p expression was increased and the TSP-1 mRNA expression was decreased in the EPCs after the intervention. miR-21-5p overexpression can improve EPCs function and inhibit TSP-1 expression but has no effect on senescence rate. Inhibition of TSP-1 expression could improve the function and reduce the senescence rate. (4) Conclusions: Our results indicate that long-term aerobic exercise can improve the functions of EPCs in aging individuals by downregulating TSP-1 expression via miR-21-5p, which reveals the mechanism of exercise in improving cardiovascular function.


Asunto(s)
Células Progenitoras Endoteliales , MicroARNs , Ratas , Masculino , Animales , Células Progenitoras Endoteliales/metabolismo , MicroARNs/genética , Trombospondina 1/metabolismo , Células Cultivadas , Ratas Sprague-Dawley
11.
Artículo en Inglés | MEDLINE | ID: mdl-36674144

RESUMEN

Maternal high-caloric nutrition and related gestational diabetes mellitus (GDM) are relevant modulators of the intrauterine environment, increasing the risk of liver metabolic alterations in mothers and offspring. In contrast, as a non-pharmacological approach against metabolic disorders, exercise is highly recommended in GDM treatment. We analysed whether gestational exercise (GE) protects mothers from diet-induced GDM metabolic consequences and mitigates liver mitochondrial deleterious alterations in their 6-week-old male offspring. Female Sprague Dawley rats were fed with control or high-fat high-sucrose (HFHS) diet and kept sedentary or submitted to GE. Male offspring were sedentary and fed with control diet. Sedentary HFHS mothers and their offspring showed impaired hepatic mitochondrial biogenesis and morphological evidence of mitochondrial remodelling. In contrast, GE-related beneficial effects were demonstrated by upregulation of mitochondrial biogenesis signalling markers and mitochondrial fusion proteins and downregulation of mitochondrial fission protein. Alterations in miR-34a, miR-130b, and miR-494, associated with epigenetic regulation of mitochondrial biogenesis, suggested that GE is a more critical modulator of intergenerational changes in miRs expression than the maternal diet. Our data showed that GE positively modulated the altered hepatic mitochondrial biogenesis and dynamics markers and quality control signalling associated with maternal HFHS-diet-related GDM in mothers and offspring.


Asunto(s)
Diabetes Gestacional , MicroARNs , Embarazo , Ratas , Humanos , Animales , Masculino , Femenino , Sacarosa/efectos adversos , Sacarosa/metabolismo , Ratas Sprague-Dawley , Epigénesis Genética , Diabetes Gestacional/inducido químicamente , Hígado/metabolismo , Dieta Alta en Grasa/efectos adversos , MicroARNs/metabolismo
12.
Comput Math Methods Med ; 2023: 6033020, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36714328

RESUMEN

Spinal cord injury (SCI) is a serious disorder of the central nervous system with a high disability rate. Long noncoding RNAs (lncRNAs) are reported to mediate many biological processes. The aim of this study was to explore lncRNA and mRNA expression profiles and functional networks after SCI. Differentially expressed genes between SCI model rats and sham controls were identified by microarray assays and analyzed by functional enrichment. Key lncRNAs were identified using a support vector machine- (SVM-) recursive feature elimination (RFE) algorithm. A trans and cis regulation model was used to analyze the regulatory relationships between lncRNAs and their targets. An lncRNA-related ceRNA network was established. We identified 5465 differentially expressed lncRNAs (DE lncRNAs) and 8366 differentially expressed mRNAs (DE mRNAs) in the SCI group compared with the sham group (fold change > 2.0, p < 0.05). Four genes were confirmed by qRT-PCR which were consistent with the microarray data. GSEA analysis showed that most marked changes occurred in pathways related to immune inflammation and nerve cell function, including cytokine-cytokine receptor interaction, neuroactive ligand-receptor interaction, and GABAergic synapse. Enrichment analysis identified 30 signaling pathways, including those associated with immune inflammation response. A total of 40 key lncRNAs were identified using the SVM-RFE algorithm. A key lncRNA-mRNAs coexpression network was generated for 230 951 lncRNA-mRNA pairs with half showing positive correlations. Several key DE lncRNAs were predicted to have "cis"- or "trans"-regulated target genes. The transcription factors, Sp1, JUN, and SOX10, may regulate the interaction between XR_001837123.1 and ETS 1. In addition, five pairs of ceRNA regulatory sequences were constructed. Many mRNAs and lncRNAs were found to be dysregulated after SCI. Bioinformatic analysis showed that DE lncRNAs may play crucial roles in SCI. It is anticipated that these findings will provide new insights into the underlying mechanisms and potential therapeutic targets for SCI.


Asunto(s)
MicroARNs , ARN Largo no Codificante , Traumatismos de la Médula Espinal , Ratas , Animales , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Traumatismos de la Médula Espinal/genética , Traumatismos de la Médula Espinal/metabolismo , Inflamación , Redes Reguladoras de Genes , Perfilación de la Expresión Génica , MicroARNs/genética
13.
Adv Clin Chem ; 112: 205-248, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36642484

RESUMEN

Congestive heart failure (CHF) is the leading cause of morbidity and mortality in the elderly worldwide. Although many biomarkers associated with in heart failure, these are generally prognostic and identify patients with moderate and severe disease. Unfortunately, the role of biomarkers in decision making for early and advanced heart failure remains largely unexplored. Previous studies suggest the natriuretic peptides have the potential to improve the diagnosis of heart failure, but they still have significant limitations related to cut-off values. Although some promising cardiac biomarkers have emerged, comprehensive data from large cohort studies is lacking. The utility of multiple biomarkers that reflect various pathophysiologic pathways are increasingly being explored in heart failure risk stratification and to diagnose disease conditions promptly and accurately. MicroRNAs serve as mediators and/or regulators of renin-angiotensin-induced cardiac remodeling by directly targeting enzymes, receptors and signaling molecules. The role of miRNA in HF diagnosis is a promising area of research and further exploration may offer both diagnostic and prognostic applications and phenotype-specific targets. In this review, we provide insight into the classification of different biochemical and molecular markers associated with CHF, examine clinical usefulness in CHF and highlight the most clinically relevant.


Asunto(s)
Insuficiencia Cardíaca , MicroARNs , Humanos , Péptido Natriurético Encefálico , Biomarcadores , Pronóstico , Insuficiencia Cardíaca/diagnóstico , Fragmentos de Péptidos
14.
Biomater Adv ; 145: 213267, 2023 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-36599197

RESUMEN

The use of gene-based products, such as DNA or RNA, is increasingly being explored for various innovative therapies. However, the success of these strategies is highly dependent on the effective delivery of these biomolecules to target cells. Therefore, the development of pH-responsive nanoparticles comprises the creation of intelligent delivery systems with high therapeutic efficiency. In this work, the pH-responsiveness of the poly(2-(diisopropylamino)ethyl methacrylate)) (PDPA) block was investigated for the encapsulation and delivery of small RNAs (sRNA) to cancer cells. The pH responsiveness was dependent on the protonation profile of the tertiary amines of PDPA, which directly affected the electrostatic interactions established with RNA. Thus, block copolymers based on poly(oligo(ethylene oxide) methyl ether methacrylate) (POEOMA) and PDPA, POEOMA-b-PDPA, were synthesized by supplemental activator and reducing agent atom transfer radical polymerization (SARA ATRP). The structure of the block copolymers was characterized by size exclusion chromatography and 1H NMR spectroscopy. The copolymers allowed effective complexation of model sRNAs and a pre-miRNA with efficiencies of about 89 % and 91 %, respectively. The characterization by dynamic light scattering revealed that these systems had sizes between 76 and 1375 nm. It was also found that the morphology of the polyplexes depended on the pH, since the preparation at a pH lower than the pKa of the copolymers resulted in spherical but polydisperse particles, while higher pH values resulted in nanoparticles with more homogeneous size, but altered morphology. Moreover, due to pH-responsiveness, it was achieved the release of RNA at pH higher than the pKa of the copolymers, while maintaining its integrity. The polyplexes also showed a high potential to protect RNA from RNases. The transfection of a lung cancer model (A549) and fibroblast cell lines showed that these polyplexes did not cause cell toxicity. In addition, the polyplexes enabled the effective transfection of the A549 cell line with pre-miRNA-29b and miRNA-29b, resulting in a decrease of expression levels of the target DNMT3B gene by approximately 51 % and 47 %, respectively. Overall, the POEOMA-b-PDPA copolymers proved to be a promising strategy for developing responsive delivery systems, that can play a critical role in some diseases, such as cancer, where pH varies between the intra and extracellular environments.


Asunto(s)
MicroARNs , Nanopartículas , Polímeros , Metacrilatos/química , Nanopartículas/química , Concentración de Iones de Hidrógeno
15.
Sci Rep ; 13(1): 126, 2023 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-36599866

RESUMEN

Periodontitis is one of the main frequent intraoral diseases. Pathogenesis triggers are the immune responses with pro-inflammatory cytokines production and non-coding RNAs expression. The purpose of the present study was to evaluate the involvement of selected miRNAs in various stages of periodontitis and their relationship with the levels of inflammatory mediators in gingival crevicular fluid (GCF). For this study, 36 subjects (21 with periodontal disease, 15 healthy controls) were selected with an age mean of 59.1 ± 3.7 years. Clinical parameters included plaque index, gingival index, sulcus bleeding index, pocket depth, and clinical attachment level. The GCF samples were taken using capillary paper. The levels of miRNAs in GCF were estimated using a Real-Time PCR and TNFα and IL-6 levels were assessed by enzyme-linked immunosorbent assay (ELISA). The results indicated that the miRNA-103a-3p, miRNA-23a-3p, miRNA-15a-5p, and miRNA-223-3p were significantly upregulated with respect to healthy controls. Significant differences were observed for miRNA-23a-3p, miRNA-103a-3p and miRNA-423-5p levels in accord with the disease stages. Inflammatory mediators evaluated in GCF correlate well with the clinical parameters and the severity of the periodontal disease. miRNAs can represent biomarkers of disease stage and can be investigated as a possible therapeutic target, as well as levels of TNFα and IL-6 may drive the disease progression by acting as prognostic markers.


Asunto(s)
MicroARNs , Periodontitis , Humanos , Persona de Mediana Edad , Biomarcadores , Interleucina-6/química , MicroARNs/química , MicroARNs/genética , MicroARNs/metabolismo , Periodontitis/diagnóstico , Periodontitis/genética , Periodontitis/metabolismo , Factor de Necrosis Tumoral alfa/química , Factor de Necrosis Tumoral alfa/genética , Pronóstico
16.
Anal Chem ; 95(2): 1490-1497, 2023 Jan 17.
Artículo en Inglés | MEDLINE | ID: mdl-36596235

RESUMEN

In this work, a high-efficiency controllable three-dimensional (3D) DNA nanomachine (CDNM) was reasonably developed by regulating the diameter of the core and the length of the DNAzyme cantilever, which acquired greater amplification efficiency and speedier walking rate than traditional 3D DNA nanomachines with gold nanoparticles as the cores and DNAzymes as the walking arms. Significantly, once the target miRNA-21 existed, a large number of silent DNAzymes on the CDNM could be activated by enzyme-free-target-recycling amplification (EFTRA) to achieve fast cleavage and walking on the biosensor surface under the interaction of Mg2+. Impressively, when the diameter of the core was 40 nm and the length of the DNAzyme cantilever was 5 nm (15 bp), the CDNM could complete the reaction process in 60 min that was at least twice shorter than those of conventional DNA nanomachines. Moreover, the designed electrochemical biosensor successfully detected target miRNA-21 at an ultrasensitive level with a wide response range (100 aM to 1 nM) and a low detection limit (33.1 aM). Therefore, the developed CDNM provides a new idea for exploring functional DNA nanomachines in the field of biosensing for applications.


Asunto(s)
Técnicas Biosensibles , ADN Catalítico , Nanopartículas del Metal , MicroARNs , MicroARNs/genética , Oro , Límite de Detección , ADN , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos
17.
Anal Chem ; 95(2): 1193-1200, 2023 Jan 17.
Artículo en Inglés | MEDLINE | ID: mdl-36602461

RESUMEN

Sensitive and specific assay of microRNAs (miRNAs) is beneficial to early disease screening. Herein, we for the first time proposed clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a-mediated photoelectrochemical biosensors for the direct assay of miRNA-21. In this study, compared with traditional nucleic acid-based signal amplification strategies, the CRISPR/Cas13a system can greatly improve the specificity and sensitivity of target determination due to its accurate recognition and high-efficient trans-cleavage capability without complex nucleic acid sequence design. Moreover, compared with the CRISPR/Cas12a-based biosensing platform, the developed CRISPR/Cas13a-mediated biosensor can directly detect RNA targets without signal transduction from RNA to DNA, thereby avoiding signal leakage and distortion. Generally, the proposed biosensor reveals excellent analysis capability with a wider linear range from 1 fM to 5 nM and a lower detection limit of 1 fM. Additionally, it also shows satisfactory stability in the detection of human serum samples and cell lysates, manifesting that it has great application prospects in the areas of early disease diagnosis and biomedical research.


Asunto(s)
Investigación Biomédica , Técnicas Biosensibles , MicroARNs , Humanos , Bioensayo , Transducción de Señal
18.
Funct Integr Genomics ; 23(1): 33, 2023 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-36625940

RESUMEN

Human diseases have been a critical threat from the beginning of human history. Knowing the origin, course of action and treatment of any disease state is essential. A microscopic approach to the molecular field is a more coherent and accurate way to explore the mechanism, progression, and therapy with the introduction and evolution of technology than a macroscopic approach. Non-coding RNAs (ncRNAs) play increasingly important roles in detecting, developing, and treating all abnormalities related to physiology, pathology, genetics, epigenetics, cancer, and developmental diseases. Noncoding RNAs are becoming increasingly crucial as powerful, multipurpose regulators of all biological processes. Parallel to this, a rising amount of scientific information has revealed links between abnormal noncoding RNA expression and human disorders. Numerous non-coding transcripts with unknown functions have been found in addition to advancements in RNA-sequencing methods. Non-coding linear RNAs come in a variety of forms, including circular RNAs with a continuous closed loop (circRNA), long non-coding RNAs (lncRNA), and microRNAs (miRNA). This comprises specific information on their biogenesis, mode of action, physiological function, and significance concerning disease (such as cancer or cardiovascular diseases and others). This study review focuses on non-coding RNA as specific biomarkers and novel therapeutic targets.


Asunto(s)
MicroARNs , Neoplasias , ARN Largo no Codificante , Humanos , ARN no Traducido/genética , MicroARNs/genética , MicroARNs/metabolismo , Biomarcadores , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , ARN Circular/genética , Neoplasias/genética , Neoplasias/terapia
19.
Int J Oncol ; 62(2)2023 02.
Artículo en Inglés | MEDLINE | ID: mdl-36633145

RESUMEN

MicroRNA (miRNA), a non­coding single­stranded RNA molecule with a length of 21­25 nucleotides transcripts, has been identified to play important roles in tumorigenesis and shows great potential applications in cancer diagnosis, prognosis and therapy. Brain derived neurotrophic factor (BDNF) is a member of the nerve growth factor family and usually serves as a biomarker in neurological and neuropsychiatric diseases for diagnosis and treatment by regulating its high­affinity receptor TrkB (Tyrosine Kinase Receptor B). Abnormal expression of BDNF is also closely related to the development of cancer, cancer­related pain and depression. However, little significant progress has been made in the application of BDNF in cancers. Recent studies have shown that the expression of BDNF is directly regulated by a cluster of miRNAs. This review concluded and discussed the role and mechanism of miRNAs targeting BDNF in cancers, and provided novel insights into the diagnosis and therapy of cancer in the future.


Asunto(s)
MicroARNs , Neoplasias , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Factor Neurotrófico Derivado del Encéfalo/genética , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Receptor trkB/genética , Receptor trkB/metabolismo , Pronóstico , Neoplasias/diagnóstico , Neoplasias/genética , Neoplasias/terapia
20.
Cell Death Dis ; 14(1): 23, 2023 Jan 13.
Artículo en Inglés | MEDLINE | ID: mdl-36635261

RESUMEN

Glioma is the most aggressive and common malignant neoplasms in human brain tumors. Numerous studies have showed that glioma stem cells (GSCs)drive the malignant progression of gliomas. Recent studies have revealed that circRNAs can maintain stemness and promote malignant progression of glioma stem cells. We used bioinformatics analysis to identify circRNAs and potential RNA-binding proteins (RBPs) in glioma. qRT-PCR, western blotting, RNA FISH, RNA pull-down, RNA immunoprecipitation assay, ChIP, immunohistochemistry, and immunofluorescence methods were used to quantified the expression of circNCAPG, U2AF65, RREB1 and TGF-ß1, and the underlying mechanisms between them. MTS, EDU, neurosphere formation, limiting dilution neurosphere formation and transwell assays examined the proliferation and invasive capability of GSCs, respectively. We identified a novel circRNA named circNCAPG was overexpressed and indicated the poor prognosis in glioma patients. Upregulating circNCAPG promoted the malignant progression of GSCs. RNA binding protein U2AF65 could stabilize circNCAPG by direct binding. Mechanically, circNCAPG interacted with and stabilized RREB1, as well as stimulated RREB1 nuclear translocation to activate TGF-ß1 signaling pathway. Furthermore, RREB1 transcriptionally upregulated U2AF65 expression to improve the stability of circNCAPG in GSCs, which established a feedback loop involving U2AF65, circNCAPG and RREB1. Since circRNA is more stable than mRNA and can execute its function continuously, targeting circNCAPG in glioma may be a novel promising therapeutic.


Asunto(s)
Neoplasias Encefálicas , Glioma , MicroARNs , ARN Circular , Humanos , Neoplasias Encefálicas/patología , Línea Celular Tumoral , Proliferación Celular/genética , Proteínas de Unión al ADN/metabolismo , Retroalimentación , Regulación Neoplásica de la Expresión Génica , Glioma/patología , MicroARNs/genética , Células Madre Neoplásicas/metabolismo , ARN Circular/genética , Factores de Transcripción/metabolismo , Factor de Crecimiento Transformador beta1/genética , Factor de Crecimiento Transformador beta1/metabolismo , Factor de Empalme U2AF/genética , Factor de Empalme U2AF/metabolismo
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