RESUMO
MicroRNAs (miRNAs) are endogenous 21-nucleotide small RNAs that direct sequence-specific silencing of complementary messenger RNAs to regulate a wide range of biological processes. In plants, miRNA precursors are processed from imperfect foldback structures by the RNase III enzyme DICER-LIKE1, in coordination with accessory proteins. While mismatches flanking the miRNA/miRNA* duplex in endogenous precursors can strongly influence miRNA accumulation, their impact has not been thoroughly examined in the context of artificial miRNAs (amiRNAs) used for targeted gene silencing in plants. Here, using silencing sensor systems in Nicotiana benthamiana, we systematically investigated how base pairing at or near DCL1 cleavage sites affects amiRNA production from the recently described minimal shc precursor. Independent pairing of naturally mismatched positions revealed that introducing a G-C pair immediately upstream of the mature amiRNA remarkably enhances amiRNA accumulation and silencing efficiency. This effect was further validated in Arabidopsis transgenic lines targeting endogenous genes and confirmed by deep sequencing, which revealed highly accurate processing and predominant release of the intended amiRNAs, supporting the specificity of the approach. Our findings show that a single structural modification in an amiRNA precursor can significantly enhance the efficacy of amiRNA-mediated gene silencing. This optimized amiRNA platform is well suited for large-scale functional genomics screens and should facilitate the development of next-generation crops with enhanced resilience to environmental stresses.
Assuntos
Inativação Gênica , MicroRNAs , Nicotiana , RNA de Plantas , MicroRNAs/genética , MicroRNAs/metabolismo , Nicotiana/genética , Nicotiana/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Pareamento de Bases , Plantas Geneticamente Modificadas , RNA de Plantas/genética , RNA de Plantas/metabolismo , Regulação da Expressão Gênica de PlantasRESUMO
Epigenetic modifications function as central controllers of gene expression in cancer, coordinating crucial cellular activities that trigger the initiation and progression of the tumor, besides their importance in therapeutic response. These modifications can control the gene expression without changing the sequence of DNA. In colorectal cancer (CRC), these alterations involving DNA methylation, histone modifications, chromatin rearrangement, and noncoding ribonucleic acids play a significant role in the pathogenesis of CRC. Abnormal DNA methylation silences the tumor suppressor genes, meanwhile leading to the instability of the genome via reduction of the whole methylation. Specific methylation signatures, such as CpG islands, help in categorizing the subtypes of the tumor and predicting the clinical outcomes. In addition, histone-modifying enzymes, including enhancer of zeste homolog 2 and histone deacetylases, are frequently uncontrolled in cancer, leading to alterations in gene expression. Moreover, small regulatory ribonucleic acids such as microRNA-21 and microRNA-143 contribute to the complex networks that regulate cell survival and growth. Collectively, these epigenetic alterations trigger the transition from benign growth to malignant cancer by continuously suppressing crucial genes. Furthermore, the epigenetic markers can be detected in blood and stool specimens, offering promising tools for the early detection of cancer. The major obstacle in cancer treatment is the resistance to chemotherapy drugs, which is mainly caused by epigenetic modifications in cancer cells. Therefore, the new therapeutic ways target the modifications that occur in DNA methylation and histone, mostly in conjunction with conventional therapies. As the metabolites produced by the gut microbiome can alter the host epigenetics, they can promote cancer development. Promising technologies help in the concise proofreading of epigenetic marks, and advanced single-cell analysis is paving the way for personalized treatment approaches. This cutting-edge knowledge of epigenetic regulation mechanisms offers new prospects for enhancing diagnosis, prognosis, and targeted therapies in colorectal cancer.
Assuntos
Neoplasias Colorretais , Metilação de DNA , Epigênese Genética , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Neoplasias Colorretais/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Biomarcadores Tumorais/genética , Medicina de PrecisãoRESUMO
BACKGROUND: Tumor metastasis is a key factor in cancer progression, yet its molecular mechanisms are not fully understood. ERBB2-positive lung cancer exhibits aggressive behavior, and the role of miR-139 in its metastasis requires investigation. OBJECTIVES: This study aimed to explore the function of miR-139 in ERBB2-positive lung cancer and its underlying molecular mechanism involving the ERBB2/Rac1/NF-κB signaling axis. METHODS: The study utilized A549 lung cancer cells and tissue samples from 106 lung cancer patients. Methods included RT-PCR, bioinformatics analysis, dual-luciferase reporter assay, Western blot, cell migration/invasion assays, wound healing tests, Rac1 activity assays, and rescue experiments using Rac1-Q61L. RESULTS: MiR-139 expression was significantly downregulated in lung cancer tissues, especially in lymph node metastases (P<0.01). MiR-139 directly targeted the 3'UTR of ERBB2 and inhibited its expression (P<0.01). Overexpression of miR-139 reduced Rac1 activity (P<0.01) without affecting RhoA or Cdc42, and decreased NF-κB signaling activity in ERBB2-positive tissues. MiR-139 overexpression significantly suppressed cell migration and invasion (P<0.01), an effect partially reversed by Rac1-Q61L. CONCLUSION: MiR-139 inhibits lung cancer cell migration and invasion by targeting ERBB2, suppressing Rac1 activity, and downregulating NF-κB signaling. Its downregulation promotes metastasis through the ERBB2/Rac1/NF-κB axis.
Assuntos
Neoplasias Pulmonares , MicroRNAs , NF-kappa B , Receptor Proteína Tirosina Quinase ErbB-2 , Proteínas rac1 de Ligação ao GTP , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Proteínas rac1 de Ligação ao GTP/metabolismo , Proteínas rac1 de Ligação ao GTP/genética , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Transdução de Sinais , Receptor Proteína Tirosina Quinase ErbB-2/metabolismo , Receptor Proteína Tirosina Quinase ErbB-2/genética , Regulação para Baixo , NF-kappa B/metabolismo , NF-kappa B/genética , Movimento Celular/genética , Regulação Neoplásica da Expressão Gênica , Células A549 , Invasividade Neoplásica , Metástase Neoplásica , Masculino , Feminino , Metástase LinfáticaRESUMO
Coronary artery disease, a leading cause of mortality, underscores the need for biomarkers that can predict acute coronary syndrome (ACS) occurrence and outcomes. MicroRNAs (miRNAs) are increasingly recognized as potential markers. This study aimed to identify a plasma miRNA signature in ACS patients undergoing elective cardiac catheterization and to investigate miR-100's potential as an ACS prognostic biomarker. Plasma samples from 100 patients with suspected ACS were analyzed. qPCR revealed significantly elevated plasma miR-29a-3p, miR-100, miR-192, and miR-194-5p in ACS patients, and multiplex ELISA showed increased myeloperoxidase. Sub-network enrichment analysis identified TP53 as a central regulator of the miRNA-gene interaction network. In H9c2 cardiomyoblasts, hypoxic treatment (1% O2) induced significant cytotoxicity and increased intracellular and released miR-100 levels. Western blotting further showed that hypoxia suppressed p53, HMGB1, NF-κB, and Bcl-2 expression. Consistent with the predicted regulatory network, siRNA-mediated p53 silencing markedly reduced constitutive miR-100 expression and triggered compensatory upregulation of inflammatory and survival-related proteins, including TLR4, NF-κB, and Bcl-2. Conversely, miR-100 overexpression significantly increased p53 protein levels and reduced the pro-survival factor Mcl-1, whereas miR-100 inhibition produced the opposite effect. These findings define a reciprocal p53/miR-100 regulatory axis that influences inflammatory and survival signaling in cardiomyoblasts. In conclusion, both clinical profiling and mechanistic studies support miR-100 as a promising early prognostic biomarker for ACS and suggest that hypoxia-induced disruption of the p53/miR-100 axis may contribute to cardiomyoblast vulnerability. Further studies are warranted to explore its therapeutic potential.
Assuntos
Síndrome Coronariana Aguda , MicroRNAs , Mioblastos Cardíacos , Proteína Supressora de Tumor p53 , MicroRNAs/sangue , MicroRNAs/metabolismo , MicroRNAs/genética , Síndrome Coronariana Aguda/diagnóstico , Síndrome Coronariana Aguda/genética , Síndrome Coronariana Aguda/sangue , Humanos , Proteína Supressora de Tumor p53/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/fisiologia , Biomarcadores/sangue , Biomarcadores/metabolismo , Masculino , Pessoa de Meia-Idade , Animais , Prognóstico , NF-kappa B/metabolismo , Feminino , Mioblastos Cardíacos/metabolismo , RatosRESUMO
Mitochondria are key players in regulating cellular metabolism. Short noncoding RNAs, or microRNAs (miRNAs), have become significant modulators of gene expression and cellular functions. Recent research has demonstrated the presence and activity of mitochondrial miRNAs (MitomiRs) in various cell types, including blood cells. The role of MitomiRNAs in metabolic reprogramming throughout blood cell formation, including their biosynthesis, function, and possible therapeutic implications, was discussed in this chapter. The discovery of mitochondrial microRNAs (MitomiRs) transformed our understanding of gene regulation in these critical organelles. These MitomiRs were discovered as they were influencing mitochondrial gene expression, with considerable effects on various cellular functions like oxidative stress responses, production of energy, and apoptosis during blood cell development.
Assuntos
Células Sanguíneas , MicroRNAs , Mitocôndrias , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Mitocôndrias/genética , Mitocôndrias/metabolismo , Estresse Oxidativo/genética , Animais , Células Sanguíneas/metabolismo , Células Sanguíneas/citologia , Regulação da Expressão Gênica , Apoptose/genética , Metabolismo Energético/genética , Reprogramação Celular , Reprogramação MetabólicaRESUMO
Small, endogenous noncoding RNAs known as miRNAs are widely distributed and use posttranscriptional control to suppress the expression of target genes. miRNAs have crucial roles in the development of disease and carcinogenesis, and new research suggests that miRNAs might be reliable diseases biomarkers. The identification of miRNAs in bodily fluids, in particular, has created a compelling possibility for the creation of noninvasive biomarkers for disease diagnosis, prognosis, and response prediction to treatment. Deregulation of the expression of microRNAs, which are essential for the regulation of several biological processes, has been connected to the emergence of metabolic, cardiovascular, neurodegenerative, and cancerous disorders. An exhaustive, up-to-date, and comprehensive investigation of the role of miRNAs in disease is presented in this chapter. Its goal is to stimulate further study in this field since small miRNAs may be utilized for effective disease detection, prognosis, and therapy.
Assuntos
Biomarcadores Tumorais , Doenças Cardiovasculares , MicroRNAs , Neoplasias , Doenças Neurodegenerativas , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias/diagnóstico , Neoplasias/genética , Prognóstico , Biomarcadores Tumorais/genética , Doenças Cardiovasculares/diagnóstico , Doenças Cardiovasculares/genética , Doenças Neurodegenerativas/diagnóstico , Doenças Neurodegenerativas/genética , Regulação Neoplásica da Expressão GênicaRESUMO
Cancer is a genomically complex and multifaceted disease. Wealth of information distilled through decades of high-throughput research has revealed a broad spectrum of oncogenic signaling cascades, immunological evasion, and drug resistance, which play a steering role in carcinogenesis and metastasis. Noncoding RNAs have also emerged as key players in the regulation of multiple stages of cancer progression and metastatic spread of cancer cells to secondary sites. The concept of probiotics has started to gain limelight due to its ability to pharmacologically modulate the host microbiome and the immunological responses. The genomics era has provided impetus for the discovery and characterization of bacterial probiotic effector molecules that stimulate specific responses. We have witnessed an exponential increase in the seminal studies which provided proof-of-concept about the mechanistic regulation of cell signaling pathways and noncoding RNAs by probiotics. These exciting and groundbreaking studies ignited an outburst of data generated using several "omics" technologies. In this chapter, we have provided a summary of seminal studies associated with the anticancer and antimetastatic role of probiotics in animal models. However, circumstantial evidence has also underlined tumor-promoting role of probiotics in animal model studies. Therefore, there is a need to scrupulously reinterpret the existing pieces of evidence related to conflicting data about pro-tumorigenic and tumor-inhibitory roles of probiotics. We also critically summarized how probiotics modulated noncoding RNAs to prevent/inhibit cancer progression. Surprisingly, probiotics-mediated regulation of noncoding RNAs has not been comprehensively explored in different cancers. In accordance with this approach, in-depth analysis of target long noncoding RNAs and circular RNAs by probiotics will allow the researchers to develop near-to-complete signaling landscape to reap the full benefits of the medicinal significance of probiotics.
Assuntos
Carcinogênese , MicroRNAs , Neoplasias , Probióticos , RNA Longo não Codificante , Probióticos/uso terapêutico , RNA Longo não Codificante/genética , Humanos , Animais , MicroRNAs/genética , Carcinogênese/genética , Carcinogênese/efeitos dos fármacos , Neoplasias/genética , Neoplasias/patologia , Neoplasias/microbiologia , Metástase Neoplásica , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacosRESUMO
In recent years, noncoding RNAs have sparked significant interest in understanding the diverse roles of ncRNAs in cellular regulation and disease processes. Noncoding RNAs (ncRNAs), encompassing small noncoding RNAs (sncRNAs) and long noncoding RNAs (lncRNAs), are critical regulators of gene expression, epigenetic modifications, and various cellular processes within the human genome. The diverse nature ncRNAs, along with certain complex features, has made them difficult to study through traditional experimental methods. As a result, bioinformatics tools have expanded the possibilities for offering new insights through advanced computational strategies. This chapter explores the recent advancements in ncRNA databases, emphasizing their importance and the innovative in silico strategies that enable the prediction and analysis of biological interactions, particularly for miRNAs and lncRNAs. It offers an in-depth overview of the structural properties, classification, and functions of various types of noncoding RNAs, highlighting their crucial roles in cellular processes. Additionally, the chapter discusses the significant therapeutic potential of ncRNAs, focusing on their applications in treating cancer and other severe diseases. These insights are pivotal in advancing the development of targeted therapies and precision medicine.
Assuntos
Biologia Computacional , MicroRNAs , Neoplasias , RNA Longo não Codificante , RNA não Traduzido , Humanos , Biologia Computacional/métodos , RNA Longo não Codificante/genética , RNA não Traduzido/genética , MicroRNAs/genética , Neoplasias/genética , Neoplasias/terapia , Epigênese GenéticaRESUMO
Breast cancer remains one of the leading causes of cancer-related death among women worldwide, characterized by significant molecular and metabolic heterogeneity. Metabolic reprogramming has been shown to enable tumor cells to adapt to the dynamic microenvironment, supporting uncontrolled proliferation and survival. Emerging evidence highlights the critical roles of noncoding RNAs (ncRNAs), including microRNAs, long noncoding RNAs, and circular RNAs, in coordinating the complex regulatory networks underlying metabolic reprogramming in breast cancer. These ncRNAs influence key metabolic pathways such as glycolysis, lipid metabolism, and amino acid metabolism by targeting transcription factors, enzymes, and signaling cascades. Evidence suggests that targeting dysregulated ncRNAs holds significant potential for modulating cancer cell metabolism and offers novel strategies for breast cancer management. Furthermore, bioactive compounds derived from dietary sources have demonstrated the ability to modulate ncRNA expression and function, presenting exciting prospects for dietary or nutritional interventions in breast cancer therapy. This chapter compiles the intricate relationship between ncRNAs and metabolic reprogramming in breast cancer, with a focus on innovative techniques to target ncRNAs and the potential of dietary strategies to influence these regulatory pathways.
Assuntos
Neoplasias da Mama , RNA não Traduzido , Humanos , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Neoplasias da Mama/dietoterapia , Neoplasias da Mama/patologia , Feminino , RNA não Traduzido/genética , RNA não Traduzido/metabolismo , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Glicólise/genética , Microambiente Tumoral , Animais , Metabolismo dos Lipídeos/genética , RNA Circular , Pesquisa Translacional Biomédica , Reprogramação MetabólicaRESUMO
The clinical potential of noncoding RNA has proven to have major advancements recently. Understanding their mechanisms and biological functions drives the use of ncRNAs in clinical applications. ncRNAs play crucial roles in regulating all cellular processes by influencing key proteins involved in the singling pathways, both at gene transcriptional and translational levels. Additionally, ncRNAs can be detected intracellularly and extracellularly in different body fluids such as blood, urine, and cerebrospinal fluid, facilitating their noninvasive use in clinical settings. Several clinical trials have investigated ncRNAs' therapeutic and diagnostic use, leading to models that mimic the mechanistic functions of dysregulated ncRNAs. These models have provided new clinical tools for early disease diagnosis and therapy. While the FDA has approved some ncRNA-based diagnostic/therapeutic applications, other clinical trials were withdrawn or terminated. Challenges associated with ncRNAs include off-target effects due to a lack of specificity, decreased stability of specific ncRNAs in vivo, low cellular uptake, and limitations in delivery systems. Additionally, most exciting studies focus primarily on some types of ncRNA, such as microRNAs, highlighting the need for broader research into investigating other types of ncRNAs. Unlocking the full potential of ncRNAs will pave the way for a myriad of possibilities for developing novel strategies for early diagnosis, disease prognosis, and targeted therapies for different human diseases.
Assuntos
Neoplasias , RNA não Traduzido , Humanos , Ensaios Clínicos como Assunto , RNA não Traduzido/genética , RNA não Traduzido/uso terapêutico , Neoplasias/diagnóstico , Neoplasias/genética , Neoplasias/terapia , MicroRNAs/genética , MicroRNAs/uso terapêutico , AnimaisRESUMO
Kidney injury is a common complication of deep hypothermic circulatory arrest (DHCA) in patients undergoing cardiac surgery. However, the role of competitive endogenous RNA (ceRNA) networks in mediating DHCA-induced kidney injury has not been fully elucidated. In the present study, we aimed to systematically analyze ceRNA networks and identify a novel lncRNA, MSTRG.16386.1, that promotes DHCA-induced kidney injury by regulating the miR-466b-5p/Sprouty RTK signaling antagonist 2 (Spry2) axis. Kidney injury induced by DHCA was confirmed using a rat model of cardiopulmonary bypass (CPB) with or without DHCA. In addition, lncRNA and mRNA sequencing of kidney tissues revealed 309 specific shared differentially expressed mRNAs (DEGs) and 439 differentially expressed lncRNAs (DELs) in the kidneys of CPB+DHCA rats compared with those of SHAM and CPB rats. Differentially expressed miRNAs (DEMs) were predicted by coexpression and binding site analysis. A ceRNA network consisting of 12 DEGs, 6 DELs, and 11 DEMs was constructed, and the top ranked RNAs were Sprouty RTK Signaling Antagonist 2 (Spry2), MSTRG.16386.1, and miR-466b-5p, which were verified using qRT-PCR. We found that MSTRG.16386.1 overexpression resulted in NRK-52E cell apoptosis, which was suppressed by the interaction with miR-466b-5p. In addition, we demonstrated that Spry2 is a key target of MSTRG.16386.1 and miR-466b-5p. Rescue experiment results revealed that the downregulated expression of Spry2 protected NRK-52E cells against apoptosis mediated by MSTRG.16386.1 overexpression or an miR-466b-5p inhibitor. Our findings provide novel insights into ceRNA regulation of the MSTRG.16386.1/miR-466b-5p/Spry2 axis in DHCA-induced kidney injury by the induction of apoptosis.
Assuntos
Injúria Renal Aguda , Parada Circulatória Induzida por Hipotermia Profunda , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas de Membrana , MicroRNAs , RNA Longo não Codificante , Animais , MicroRNAs/genética , MicroRNAs/metabolismo , Ratos , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Masculino , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/genética , Injúria Renal Aguda/patologia , Ratos Sprague-Dawley , Parada Circulatória Induzida por Hipotermia Profunda/efeitos adversos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Transdução de Sinais , Humanos , Apoptose , RNA Endógeno Competitivo , Proteínas do Tecido NervosoRESUMO
Chronic obstructive pulmonary disease (COPD) is a common chronic respiratory disorder. Its prevention and management have become increasingly challenging owing to subtle and easily overlooked early symptoms, warranting the exploration of detection markers. Hence, this study aimed to explore COPD-related competitive endogenous RNA (ceRNA) mechanisms with notable predictive value by leveraging bioinformatics methods. Herein, Gene Expression Omnibus, lncRNASNP2, and miRDB were used to construct a COPD-related ceRNA network, which was then assessed through Kyoto Encyclopedia of Gene and Genomes pathway analysis. The targeting relationship of identified axis was confirmed through RNA pull-down and dual-luciferase reporter assays. In total, 106 patients with stable COPD and 98 patients with acute exacerbation COPD (AECOPD) were enrolled. Binary logistic regression and receiver operating characteristic curve analyses were performed to evaluate the clinical significance of the identified axis. Additionally, its role in COPD-related inflammation was investigated in BEAS-2B cells treated with cigarette smoke extract (CSE). A potential ceRNA mechanism was identified involving LINC01569, miR-4722-5p, and RAB14, where LINC01569 overexpression promoted RAB14 expression by competitively binding to miR-4722-5p. Both LINC01569 and RAB14 were highly expressed in AECOPD, making them risk factors of COPD (odds ratio [OR] = 5.100 and 8.076, respectively), with their area under the curve (AUC) values for predicting disease progression being 0.878 and 0.822, respectively. In contrast, miR-4722-5p acted as a potential protective factor (OR = 0.448), with an AUC of 0.724 for predicting AECOPD occurrence. Their serum expression strongly correlated with inflammatory markers. In CSE-treated BEAS-2B cells, silencing LINC01569 upregulated miR-4722-5p, thereby suppressing RAB14 expression and reducing pro-inflammatory factor production. Altogether, the LINC01569/miR-4722-5p/RAB14 regulatory axis represents a potential ceRNA mechanism influencing COPD progression. It demonstrates significant predictive value for disease development and plays a crucial role in COPD-associated inflammatory processes.
Assuntos
Inflamação , MicroRNAs , Doença Pulmonar Obstrutiva Crônica , RNA Longo não Codificante , Proteínas rab de Ligação ao GTP , Humanos , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/genética , Doença Pulmonar Obstrutiva Crônica/patologia , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Proteínas rab de Ligação ao GTP/genética , Proteínas rab de Ligação ao GTP/metabolismo , Masculino , Feminino , Inflamação/metabolismo , Inflamação/genética , Inflamação/patologia , Pessoa de Meia-Idade , Idoso , Linhagem CelularRESUMO
Background: Wilson's disease (WD), caused by mutations in the ATP7B gene, leads to copper accumulation and multi-organ damage. Exosomal microRNAs (miRNAs) play a crucial role in cell-to-cell communication and the pathogenesis of diseases, yet their study in WD remains unreported. This study aims to characterize the serum exosomal miRNA signature in WD patients and investigate its potential as a source of biomarkers and therapeutic targets. Methods: Serum exosomes from WD patients and healthy controls were isolated for RNA sequencing to identify differentially expressed miRNAs (DE-miRNAs). An integrated bioinformatics approach was employed, encompassing Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Reactome, and Disease Ontology (DO) analyses to systematically decipher the functional roles, pathway involvements, and disease associations of the DE-miRNAs. Selected DE-miRNAs were validated by RT-qPCR. Results: We identified 59 DE-miRNAs (23 upregulated, 34 downregulated) in WD patient serum exosomes. GO analysis revealed their significant involvement in signal transduction, metal ion binding, and metabolic pathways. KEGG analysis highlighted alterations in key signaling cascades, including Ras, PI3K-Akt, and Hippo pathways. Reactome analysis further uncovered disruptions in specific biological modules, notably ubiquitin-mediated proteolysis, GPCR signaling, and spliceosome assembly. DO enrichment demonstrated significant associations with hepatocellular carcinoma, neuropsychiatric disorders, and metabolic diseases. RT-qPCR validation confirmed the reliability of DE-miRNA expression patterns (p < 0.05). Conclusions: This study establishes the first comprehensive landscape of serum exosomal miRNAs in WD, revealing their involvement in an interconnected network of pathological processes. Our findings provide a novel conceptual framework for understanding WD pathophysiology and pinpoint promising candidates for biomarker development.
Assuntos
Biomarcadores , Exossomos , Degeneração Hepatolenticular , MicroRNAs , Humanos , Degeneração Hepatolenticular/sangue , Degeneração Hepatolenticular/genética , Degeneração Hepatolenticular/metabolismo , Exossomos/metabolismo , MicroRNAs/sangue , MicroRNAs/genética , MicroRNAs/metabolismo , Biomarcadores/sangue , Feminino , Masculino , Adulto , Transdução de Sinais , Biologia Computacional , Ontologia Genética , Adulto JovemRESUMO
Prostate cancer is the most common malignancy among men worldwide, and treatment response depends on tumor radiosensitivity. Micrornas (mirnas) are key regulators of cell proliferation, apoptosis, and dna damage response, and have been implicated in therapy resistance. However, their roles in prostate cancer radioresponse remain incompletely understood. This study investigated the expression patterns of ten selected mirnas associated with radiation resistance in other cancers in prostate cancer models. Radiation-resistant pc-3, radiation-sensitive lncap, and normal prostate epithelial (hprec) cells were exposed to 0, 2, 4, 6, and 8 gy of ionizing radiation. Mirna expression levels were analyzed by quantitative pcr using snord48 as an internal control and calculated with the 2 - δδct method. In pc-3 cells, mir-20a-5p, mir-128-3p, and mir-135b-5p showed significant dose-dependent upregulation, whereas mir-23b-3p and mir-381-3p were downregulated. Mir-128-3p correlated positively with radiation dose, while mir-23b-3p and mir-381-3p showed negative correlations. Lncap cells exhibited moderate, non-dose-dependent mirna changes. Distinct mirna signatures differentiate radiation-resistant and radiation-sensitive prostate cancer cells. Mir-20a-5p, mir-128-3p, and mir-135b-5p may contribute to radioresistance, whereas mir-23b-3p and mir-381-3p may act as radiosensitizers.
Assuntos
Regulação Neoplásica da Expressão Gênica , MicroRNAs , Neoplasias da Próstata , Tolerância a Radiação , Humanos , Masculino , MicroRNAs/genética , Neoplasias da Próstata/genética , Neoplasias da Próstata/radioterapia , Tolerância a Radiação/genética , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Linhagem Celular Tumoral , Perfilamento da Expressão Gênica , Células PC-3RESUMO
This study evaluated the clinical significance of miR-567 in atherosclerosis and its regulatory effect on PDGF-BB-induced VSMCs, aiming to identify a novel biomarker for the risk and progression of atherosclerosis. The study enrolled 113 atherosclerosis patients and 126 non-atherosclerosis patients. Serum miR-567 level was compared between the two groups and its significance in disease severity was assessed. T/G human VSMC was induced with PDGF-BB, and based on this cell model, the regulatory effect and potential mechanism of miR-567 was estimated. miR-567 was upregulated in atherosclerosis patients and showed diagnostic significance (AUC = 0.875). miR-567 was positively correlated with homocysteine, total cholesterol, and low-density lipoprotein, and negatively correlated with high-density lipoprotein (r > 0.7, P < 0.0001). miR-567 was upregulated in PDGF-BB-induced VSMCs, and silencing miR-567 showed protective effect on PDGF-BB-induced VSMCs. CSF1R was negatively correlated with miR-567. Silencing CSF1R reversed the protective effect of miR-567 silencing on PDGF-BB-induced VSMCs.
Assuntos
Aterosclerose , Becaplermina , Inflamação , MicroRNAs , Músculo Liso Vascular , Miócitos de Músculo Liso , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , MicroRNAs/sangue , Becaplermina/farmacologia , Aterosclerose/genética , Aterosclerose/patologia , Aterosclerose/metabolismo , Movimento Celular/genética , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Masculino , Feminino , Pessoa de Meia-Idade , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/patologia , Inflamação/genética , Inflamação/patologia , Fenótipo , Idoso , Proteínas Proto-Oncogênicas c-sis/farmacologia , Células CultivadasRESUMO
BACKGROUND: Long non-coding RNAs (lncRNAs) act as competing endogenous RNAs (ceRNAs) to regulate tumor progression, yet the role of LINC00536 in breast cancer (BC) remains unclear. This study investigates the oncogenic function,potential tumor microenvironment (TME) relevance, and molecular mechanism of LINC00536 in BC. METHODS: A pan-cancer analysis using TCGA/GTEx databases evaluated LINC00536 expression and its prognostic relevance. Functional assays (CCK-8, EdU, wound healing, colony formation) and qRT-PCR were performed in BC cell lines. Dual-luciferase reporter, Western blot, and ELISA validated the LINC005iR-204-5p/TGFBR2 axis and TGF-ß pathway activation. Nude mouse xenograft and lung metastasis models assessed in vivo tumorigenicity. RESULTS: LINC00536 was overexpressed in BC tissues/cell lines and its expression correlated with a poor prognosis. High LINC00536 levels were associated with elevated stromal/immune scores and immune checkpoint gene expression, suggesting a potential role in TME modulation. Mechanistically, LINC00536 sponged miR-204-5p, thereby upregulating TGFBR2, which activated TGF-ß signaling and induced epithelial-mesenchymal transition (EMT). In vitro, LINC00536 promoted proliferation, migration, and EMT, while in vivo, it accelerated tumor growth and lung metastasis (p < 0.05). Time-dependent TGF-ß activation dynamics confirmed pathway regulation via the LINC00536/miR-204-5p/TGFBR2 axis. CONCLUSIONS: LINC00536 promotes BC progression and activates TGF-ß signaling via miR-204-5p/TGFBR2, with its expression correlating with features of the TME, establishing its potential as a diagnostic/prognostic biomarker and therapeutic target.
Assuntos
Neoplasias da Mama , MicroRNAs , RNA Longo não Codificante , Receptor do Fator de Crescimento Transformador beta Tipo II , Humanos , Neoplasias da Mama/genética , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/patologia , Neoplasias da Mama/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Feminino , MicroRNAs/genética , MicroRNAs/metabolismo , Receptor do Fator de Crescimento Transformador beta Tipo II/genética , Receptor do Fator de Crescimento Transformador beta Tipo II/metabolismo , Animais , Camundongos , Linhagem Celular Tumoral , Camundongos Nus , Regulação Neoplásica da Expressão Gênica , Microambiente Tumoral , Proliferação de Células , Prognóstico , Camundongos Endogâmicos BALB C , Transição Epitelial-Mesenquimal/genética , Fator de Crescimento Transformador beta/metabolismo , RNA Endógeno CompetitivoRESUMO
We developed a highly sensitive electrochemical sensor based on zirconium-iron metal-organic frameworks (ZrFe-MOF) for the precise detection of ultra-low abundance miRNA-21. The construction of this sensor, through an innovative thermal alkaline activation strategy, exposed abundant binding sites on the surface of ZrFe-MOF, significantly improving aptamer (Apt) loading efficiency, thereby, achieving unprecedented sensitivity with a wide linear range (100 aM-10 nM, R² = 0.997) and a record-low detection limit of 19.33 aM - outperforming existing methods by 1-2 orders of magnitude. Besides, the biosensor successfully discriminates miRNA-21 expression profiles in breast cancer cell lines and breast adenocarcinoma (MDA-MB-231 vs. MCF-7 vs. MCF-10 A), showing high consistency with qPCR results. This work not only demonstrates a novel binding-site engineering strategy for nucleic acid detection but also presents a clinically viable platform for early cancer diagnosis with superior sensitivity, specificity, and reproducibility. This thermal-alkaline activation strategy could be extended to other bimetallic MOFs for multiplex miRNA detection, with future efforts targeting sensor miniaturization for point-of-care use. Current challenges include balancing the material's electron transfer efficiency with long-term storage stability and verifying the sensor's performance in large-scale clinical cohort samples.
Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Estruturas Metalorgânicas , MicroRNAs , Zircônio , MicroRNAs/análise , MicroRNAs/genética , Humanos , Técnicas Biossensoriais/métodos , Estruturas Metalorgânicas/química , Zircônio/química , Técnicas Eletroquímicas/métodos , Aptâmeros de Nucleotídeos/química , Limite de Detecção , Ferro/química , Linhagem Celular Tumoral , TemperaturaRESUMO
The present study evaluated the role of microRNA (miR)205 as a dual regulator of angiogenesis, exhibiting both proangiogenic and antiangiogenic effects depending on the biological context. miRs are small noncoding sequences that regulate gene expression at the posttranscriptional level and can be transported in extracellular vesicles (EVs), allowing them to modulate biological processes remotely. miR205 is involved in multiple cellular processes, such as proliferation, migration, apoptosis and angiogenesis. In angiogenesis its function is contradictory: On one hand, it can inhibit blood vessel formation by suppressing proangiogenic factors such as VEGF and ANG2, as demonstrated in diseases such as psoriasis, thyroid cancer and diabetic retinopathy. However, in other contexts, miR205 promotes angiogenesis by inhibiting antiangiogenic genes such as PTEN and HITT, facilitating the activation of the PI3K/AKT pathway and cell proliferation in ovarian cancer and thrombosis. Additionally, the present study highlighted the role of EVs in transferring miR205 between cells, thereby influencing angiogenesis and disease progression. Studies in myocardial infarction and cancer models have demonstrated that EVs enriched in miR205 can affect blood vessel formation and tumor progression. Similarly, in ocular diseases such as macular degeneration and diabetic retinopathy, miR205 encapsulated in EVs has shown therapeutic potential by regulating VEGF levels. In conclusion, miR205 emerges as a promising therapeutic target for angiogenic diseases. Its application in EVbased therapy could represent an innovative strategy for treating vascular disorders. However, further studies are needed to fully understand its mechanisms of action and optimize its clinical application.
Assuntos
MicroRNAs , Neovascularização Patológica , Neovascularização Fisiológica , MicroRNAs/genética , MicroRNAs/metabolismo , Humanos , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Animais , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/genética , Neovascularização Fisiológica/genética , Proliferação de Células , AngiogêneseRESUMO
Drug resistance hinders cancer treatment and directly affects cancer outcome and survival rate. While drug resistance is one of the worst features of cancers, there is limited knowledge about its mechanism and initiation in cancers. Noncoding RNAs are one of the most impactful epigenetic elements that are deregulated in many diseases. There is a strong correlation between cancers and noncoding RNAs. In this regard, we tried to find the connection between cancers and this epigenetic factor. In our investigation, we find an axis in which circular RNA, a dominant type of noncoding RNA, could regulate miRNA to control mRNA degradation. We encountered many deregulated pathways and tried to explain the mechanism underlying the CirRNA/miRNA/mRNA axis in drug resistance of cancer, including the most widely recognized resistance type of cancer in the body.
Assuntos
Resistencia a Medicamentos Antineoplásicos , MicroRNAs , Neoplasias , RNA Circular , RNA Mensageiro , Transcrição Gênica , Humanos , Resistencia a Medicamentos Antineoplásicos/genética , Epigênese Genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias/genética , Neoplasias/tratamento farmacológico , Neoplasias/patologia , RNA Circular/genética , RNA Circular/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
In the aging HIV (human immunodeficiency virus)-infected population, comorbid diseases are important determinants of morbidity and mortality. People living with HIV (PLWH) demonstrate increased lung inflammation and incidence of chronic obstructive pulmonary disease (COPD), even after adjusting for smoking status. Disruption of the lung molecular clock has been implicated in the increased lung inflammation observed in COPD and smokers. We hypothesize that the expression of HIV TAT protein in the lungs of PLWHpromotes lung inflammation and features of emphysema due to dysregulation of lung circadian rhythm, determined by the lung molecular clock genes. We demonstrate that HIV TAT upregulates miR-126-3p in primary bronchial epithelial cells and suppresses Sirtuin 1 (SIRT1), resulting in downstream effects on core circadian genes such as BMAL1 and PER2, leading to dysregulation of the lung molecular clock. This study identifies TAT/miR-126-3p/SIRT1 axis as an important mediator of HIV-induced lung inflammation in primary human bronchial epithelial cells, SPC-TAT transgenic mice with lung-specific TAT expression, and lungs from HIV-positive donors. Using single-cell RNA sequencing of lungs from 4-month-old SP-C TAT mice, we further show that these mice already exhibit significant alterations in clock gene expression and elevated expression of proinflammatory markers in their young adult stage. Understanding the pathophysiological mechanisms by which HIV disrupts the lung molecular clock and promotes inflammation may help identify therapeutic strategies to mitigate HIV-associated COPD.