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Circulating endothelial progenitor cells (EPCs) play an important role in the repair processes of damaged vessels, favoring re-endothelization of stented vessels to minimize restenosis. EPCs number and function is diminished in patients with type 2 diabetes, a known risk factor for restenosis. Considering the impact of EPCs in vascular injury repair, we conducted a meta-analysis of microarray to assess the transcriptomic profile and determine target genes during the differentiation process of EPCs into mature ECs. Five microarray datasets, including 13 EPC and 12 EC samples were analyzed, using the online tool ExpressAnalyst. Differentially expressed genes (DEGs) analysis was done by Limma method, with an | log2FC| > 1 and FDR < 0.05. Combined p-value by Fisher exact method was computed for the intersection of datasets. There were 3,267 DEGs, 1,539 up-regulated and 1,728 down-regulated in EPCs, with 407 common DEGs in at least four datasets. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed enrichment for terms related to "AGE-RAGE signaling pathway in diabetic complications." Intersection of common DEGs, KEGG pathways genes and genes in protein-protein interaction network (PPI) identified four key genes, two up-regulated (IL1B and STAT5A) and two down-regulated (IL6 and MAPK11). MicroRNA enrichment analysis of common DEGs depicted five hub microRNA targeting 175 DEGs, including STAT5A, IL6 and MAPK11, with hsa-miR-124 as common regulator. This group of genes and microRNAs could serve as biomarkers of EPCs differentiation during coronary stenting as well as potential therapeutic targets to improve stent re-endothelization, especially in diabetic patients.
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Breast cancer is a complex, heterogeneous disease at the phenotypic and molecular level. In particular, the transcriptional regulatory programs are known to be significantly affected and such transcriptional alterations are able to capture some of the heterogeneity of the disease, leading to the emergence of breast cancer molecular subtypes. Recently, it has been found that network biology approaches to decipher such abnormal gene regulation programs, for instance by means of gene co-expression networks, have been able to recapitulate the differences between breast cancer subtypes providing elements to further understand their functional origins and consequences. Network biology approaches may be extended to include other co-expression patterns, like those found between genes and non-coding transcripts such as microRNAs (miRs). As is known, miRs play relevant roles in the establishment of normal and anomalous transcription processes. Commodore miRs (cdre-miRs) have been defined as miRs that, based on their connectivity and redundancy in co-expression networks, are potential control elements of biological functions. In this work, we reconstructed miR-gene co-expression networks for each breast cancer molecular subtype, from high throughput data in 424 samples from the Cancer Genome Atlas consortium. We identified cdre-miRs in three out of four molecular subtypes. We found that in each subtype, each cdre-miR was linked to a different set of associated genes, as well as a different set of associated biological functions. We used a systematic literature validation strategy, and identified that the associated biological functions to these cdre-miRs are hallmarks of cancer such as angiogenesis, cell adhesion, cell cycle and regulation of apoptosis. The relevance of such cdre-miRs as actionable molecular targets in breast cancer is still to be determined from functional studies.
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MicroRNAs (miRNAs) are important post-transcriptional regulators of gene expression being involved in many different biological processes and play a key role in developmental timing. Additionally, recent studies have shown that miRNAs released from parasites are capable of regulating the expression of host genes. In the present work, we studied the expression patterns of ncRNAs of various intra-mammalian life-cycle stages of the liver fluke, Fasciola hepatica, as well as those packaged into extracellular vesicles and shed by the adult fluke. The miRNA expression profile of the intra-mammalian stages shows important variations, despite a set of predominant miRNAs that are highly expressed across all stages. No substantial variations in miRNA expression between dormant and activated metacercariae were detected, suggesting that they might not be central players in regulating fluke gene expression during this crucial step in the invasion of the definitive host. We generated a curated pipeline for the prediction of putative target genes that reports only sites conserved between three different prediction approaches. This pipeline was tested against an iso-seq curated database of the 3' UTR regions of F. hepatica genes to detect miRNA regulation networks within liver fluke. Several functions related to the host immune response or modulation were enriched among the targets of the most highly expressed parasite miRNAs, stressing that they might be key players during the establishment and maintenance of infection. Additionally, we detected fragments derived from the processing of tRNAs, in all developmental stages analyzed, and documented the presence of novel long tRNA fragments enriched in vesicles. We confirmed the presence of at least 5 putative vault RNAs (vtRNAs), that are expressed across different stages and enriched in vesicles. The presence of tRNA fragments and vtRNAs in vesicles raise the possibility that they could be involved in the host-parasite interaction.
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Vesículas Extracelulares , Fasciola hepatica , MicroRNAs , Animais , Fasciola hepatica/genética , Interações Hospedeiro-Parasita/genética , Mamíferos/genética , MicroRNAs/genéticaRESUMO
In many ways, cancer cells are different from healthy cells. A lot of tactical nano-based drug delivery systems are based on the difference between cancer and healthy cells. Currently, nanotechnology-based delivery systems are the most promising tool to deliver DNA-based products to cancer cells. This review aims to highlight the latest development in the lipids and polymeric nanocarrier for siRNA delivery to the cancer cells. It also provides the necessary information about siRNA development and its mechanism of action. Overall, this review gives us a clear picture of lipid and polymer-based drug delivery systems, which in the future could form the base to translate the basic siRNA biology into siRNA-based cancer therapies.
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Rheumatoid Arthritis (RA) is an autoimmune disease with unknown etiology and a global incidence around 1%, a positive family history increases the risk of RA roughly three to five times. Pain is one of the first symptoms to appear in this disease. MicroRNAs (miRNAs) belong to the class of small non-coding RNAs; they regulate multiple cellular processes including embryonic development, cellular proliferation, differentiation and apoptosis among others. A great deal of evidence points to the employment of miRNAs as therapeutic targets and biomarkers for several pathologies. The main objective of this Review is to assess how miRNAs participate in the pathogenesis of RA. Two advanced searches were conducted in databases, one using "micro-RNA" and "rheumatoid arthritis" as key words, and another one with "micro-RNA", "pain" and "nociception". In this Review, we describe how six miRNAs: miR-16-5p, miR-23b-3b, miR-124-3p, miR-146a-5p, miR-155-5p and miR-223-3p, involved in the modulation and transmission of the nociceptive input are unregulated in RA patients. Key molecular pathways involved in nociception, inflammation and autoimmune responses, are regulated by these miRNAs; the NF-κB, TNF-α, interleukins and TLR4. By means of gene repression, the miRNAs here described modulate the nociceptive process as well as the autoimmune response that characterize this disease.
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Artrite Reumatoide/metabolismo , Regulação da Expressão Gênica , MicroRNAs/biossíntese , Nociceptividade , Artrite Reumatoide/patologia , Biomarcadores/metabolismo , HumanosRESUMO
The presence of small tooth-like indentations, or serrations, characterizes leaf margins of Arabidopsis thaliana plants. The NAC family member CUP-SHAPED COTYLEDON 2 (CUC2), which undergoes post-transcriptional gene silencing by three micro-RNA genes (MIR164A, B and C), controls the extension of leaf serration. Here, we analyzed the role of AtHB1, a transcription factor (TF) belonging to the homeodomain-leucine zipper subfamily I, in shaping leaf margins. Using mutants with an impaired silencing pathway as background, we obtained transgenic plants expressing AtHB1 over 100 times compared to controls. These plants presented an atypical developmental phenotype characterized by leaves with deep serration. Transcript measurements revealed that CUC2 expression was induced in plants overexpressing AtHB1 and repressed in athb1 mutants, indicating a positive regulation exerted by this TF. Moreover, molecular analyses of AtHB1 overexpressing and mutant plants revealed that AtHB1 represses MIR164 transcription. We found that overexpression of MIR164B was able to reverse the serration phenotype of plants overexpressing AtHB1. Finally, chromatin immunoprecipitation assays revealed that AtHB1 was able to bind in vivo the promoter regions of all three MIR164 encoding loci. Altogether, our results indicate that AtHB1 directly represses MIR164 expression to enhance leaf serration by increasing CUC2 levels.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , MicroRNAs/metabolismo , Folhas de Planta/crescimento & desenvolvimento , Fatores de Transcrição/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , MicroRNAs/genética , Fenótipo , Plantas Geneticamente Modificadas , Fatores de Transcrição/genética , TranscriptomaRESUMO
A biópsia líquida é uma ferramenta diagnóstica e prognóstica já aplicada em diversos estudos com pacientes oncológicos humanos, apresentando potencial para aplicação em oncologia veterinária. Ela se baseia na detecção de produtos tumorais variados na circulação e apresenta a vantagem de ser realizada a partir de amostra de sangue e de ser pouco invasiva ao paciente, permitindo a análise do tumor em tempo real e complementando as informações da biópsia tecidual. Nesta revisão, são abordados os conceitos gerais de biópsia líquida, suas diferentes metodologias e os diferentes produtos tumorais, incluindo células tumorais circulantes, ácidos nucleicos, englobando o DNA, micro-RNA e RNA mensageiro tumorais e exossomos. São abordadas as utilidades da biópsia líquida como ferramenta prognóstica, diagnóstica, preditiva e de direcionamento de tratamento já aplicadas em estudos de medicina humana, bem como as limitações e desafios à sua implementação em larga escala. A biópsia líquida é uma ferramenta pouco conhecida em medicina veterinária até o momento, com escassos estudos publicados.
Liquid biopsy is a diagnostic and prognostic tool already reported in several studies with human oncologic patients, and shows potential for application in veterinary oncology. However, liquid biopsy is not a widely known technique in veterinary medicine, and related research is sparse. Liquid biopsy is based on the analysis of blood samples for detection of various tumoral products in circulation. It is a non-invasive technique, and provides results in real time. Information obtained from liquid biopsies can complement the information obtained from the analysis of tissue biopsy. In this review of literature, we present the background principles of liquid biopsy, its methodology, and the tumoral products that can currently be detected with this tool. In addition to circulating tumor cells, liquid biopsies allow detection of nucleic acids, including tumor DNA, micro-RNA, messenger RNA and exosomes. We present the value of liquid biopsy as a diagnostic and prognostic tool, its predictive value in tumor progression and treatment success, and usefulness to assist treatment choice. We discuss its limitations, and the challenges to implement its use in a large scale.
La biopsia líquida es una herramienta diagnóstica y pronóstica que ya ha sido aplicada en varios estudios oncológicos humanos, con potenciales aplicaciones en oncología veterinaria. Esta técnica se fundamenta en la detección de varios tipos de productos tumorales circulantes, con la ventaja de que puede ser realizada a partir de una muestra de sangre, además de ser poco invasiva para el paciente, permitiendo el análisis del tumor en tiempo real y que sirve como complemento para las informaciones obtenidas en la biopsia de tejidos convencional. Esta revisión aborda conceptos generales de la biopsia líquida, sus diferentes métodos y los productos que pueden ser detectados, incluyendo células circulantes, ácidos nucleicos (ADN, micro ADN y ARN mensajero tumorales) y exosomas. También se discuten la utilidad de la biopsia líquida como herramienta pronostica, diagnóstica, predictiva y como información para direccionar el tratamiento de acuerdo a la experiencia en estudios de medicina humana, así como también las limitaciones y desafíos de su implementación a gran escala. La biopsia líquida es una herramienta poco conocida en medicina veterinaria, con pocos estudios publicados.
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Animais , Biópsia/métodos , Biópsia/veterinária , Células Neoplásicas Circulantes , DNA de Neoplasias/sangue , Exossomos , RNA Mensageiro , Técnicas e Procedimentos Diagnósticos/veterináriaRESUMO
A biópsia líquida é uma ferramenta diagnóstica e prognóstica já aplicada em diversos estudos com pacientes oncológicos humanos, apresentando potencial para aplicação em oncologia veterinária. Ela se baseia na detecção de produtos tumorais variados na circulação e apresenta a vantagem de ser realizada a partir de amostra de sangue e de ser pouco invasiva ao paciente, permitindo a análise do tumor em tempo real e complementando as informações da biópsia tecidual. Nesta revisão, são abordados os conceitos gerais de biópsia líquida, suas diferentes metodologias e os diferentes produtos tumorais, incluindo células tumorais circulantes, ácidos nucleicos, englobando o DNA, micro-RNA e RNA mensageiro tumorais e exossomos. São abordadas as utilidades da biópsia líquida como ferramenta prognóstica, diagnóstica, preditiva e de direcionamento de tratamento já aplicadas em estudos de medicina humana, bem como as limitações e desafios à sua implementação em larga escala. A biópsia líquida é uma ferramenta pouco conhecida em medicina veterinária até o momento, com escassos estudos publicados.(AU)
Liquid biopsy is a diagnostic and prognostic tool already reported in several studies with human oncologic patients, and shows potential for application in veterinary oncology. However, liquid biopsy is not a widely known technique in veterinary medicine, and related research is sparse. Liquid biopsy is based on the analysis of blood samples for detection of various tumoral products in circulation. It is a non-invasive technique, and provides results in real time. Information obtained from liquid biopsies can complement the information obtained from the analysis of tissue biopsy. In this review of literature, we present the background principles of liquid biopsy, its methodology, and the tumoral products that can currently be detected with this tool. In addition to circulating tumor cells, liquid biopsies allow detection of nucleic acids, including tumor DNA, micro-RNA, messenger RNA and exosomes. We present the value of liquid biopsy as a diagnostic and prognostic tool, its predictive value in tumor progression and treatment success, and usefulness to assist treatment choice. We discuss its limitations, and the challenges to implement its use in a large scale.(AU)
La biopsia líquida es una herramienta diagnóstica y pronóstica que ya ha sido aplicada en varios estudios oncológicos humanos, con potenciales aplicaciones en oncología veterinaria. Esta técnica se fundamenta en la detección de varios tipos de productos tumorales circulantes, con la ventaja de que puede ser realizada a partir de una muestra de sangre, además de ser poco invasiva para el paciente, permitiendo el análisis del tumor en tiempo real y que sirve como complemento para las informaciones obtenidas en la biopsia de tejidos convencional. Esta revisión aborda conceptos generales de la biopsia líquida, sus diferentes métodos y los productos que pueden ser detectados, incluyendo células circulantes, ácidos nucleicos (ADN, micro ADN y ARN mensajero tumorales) y exosomas. También se discuten la utilidad de la biopsia líquida como herramienta pronostica, diagnóstica, predictiva y como información para direccionar el tratamiento de acuerdo a la experiencia en estudios de medicina humana, así como también las limitaciones y desafíos de su implementación a gran escala. La biopsia líquida es una herramienta poco conocida en medicina veterinaria, con pocos estudios publicados.(AU)
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Animais , Biópsia/métodos , Biópsia/veterinária , Células Neoplásicas Circulantes , DNA de Neoplasias/sangue , RNA Mensageiro , Exossomos , Técnicas e Procedimentos Diagnósticos/veterináriaRESUMO
Resumen: La insuficiencia cardiaca (IC) es una enfermedad de alto impacto que afecta en gran medida a todas las poblaciones humanas, por lo que se ha hecho necesario el desarrollo de nuevas estrategias y métodos para manejarla. Entre estas encontramos a los microRNA, pequeños RNA no codificantes que regulan la expresión genética y que aparecen como una importante opción en el diagnóstico, pronóstico y tratamiento de esta patología. Resultados de múltiples investigaciones han establecido miRNA específicos como notorios biomarcadores de la IC, puesto que estos pueden ser aislados en fluidos corporales como la sangre y la cuantificación de sus niveles se puede correlacionar con la presencia, estado o características específicas de la enfermedad. Desde el punto de la terapéutica, por su importante rol en el control de la expresión génica y la homeostasis celular, se ha explorado su uso en la prevención o tratamiento de las características patológicas de la IC. Por ello, en esta revisión se busca mostrar la importancia de la investigación biomédica en el uso de miRNA como método para el manejo de la IC, mostrando la afectación de enfermedad en el mundo, aspectos importantes sobre la biología de los miRNA, así como avances en su uso como biomarcadores y dianas terapéuticas.
Abstract: Heart failure (HF) is a high impact disease that affects all human populations, demanding the development of new strategies and methods to manage this pathology. That's why microRNAs, small noncoding RNAs that regulate gene expression, appear as an important option in the diagnosis, prognosis and treatment of this disease. MiRNAs seems to have a future on HF handling, because can be isolated from body fluids such as blood, and changes in its levels can be associated with the presence, stage and specific disease features, which makes them an interesting option as biomarkers. Also, due to the important role of these molecules on regulation of gene expression and cell homeostasis, it has been explored its potential use as a therapeutic method to prevent or treat HF. That is why this review seeks to show the importance of biomedical research involving the use of miRNAs as a method to approach the HF, showing the impact of disease in the world, aspects of miRNAs biology, and their use as biomarkers and as important therapeutic targets.
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Heart failure (HF) is a high impact disease that affects all human populations, demanding the development of new strategies and methods to manage this pathology. That's why microRNAs, small noncoding RNAs that regulate gene expression, appear as an important option in the diagnosis, prognosis and treatment of this disease. MiRNAs seems to have a future on HF handling, because can be isolated from body fluids such as blood, and changes in its levels can be associated with the presence, stage and specific disease features, which makes them an interesting option as biomarkers. Also, due to the important role of these molecules on regulation of gene expression and cell homeostasis, it has been explored its potential use as a therapeutic method to prevent or treat HF. That is why this review seeks to show the importance of biomedical research involving the use of miRNAs as a method to approach the HF, showing the impact of disease in the world, aspects of miRNAs biology, and their use as biomarkers and as important therapeutic targets.
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Pancreatic cancer (PC) is an aggressive malady with proclivity for early metastasis. Overexpression of toll-like receptor 4 (TLR4) in pancreatic ductal adenocarcinoma, the most common type of pancreatic malignancy, correlates to tumor size, lymph node involvement, venous invasion and pathological stage. Lipopolysaccharides (LPS) are natural TLR4 ligands that have been shown to increase the invasive ability of PC cells. However, rapid inactivation of circulating LPS and low systemic absorption of inhaled LPS from the bronchoalveolar compartment make other agonists such as saturated fatty acids more suitable to be considered for TLR4-related cell invasiveness. Interestingly, PC risk was strongly associated to intake of saturated fat from animal food sources, in particular to consumption of saturated palmitic acid (PA). In the present study, we investigated the influence of PA on the invasive capacity of human PC cells AsPC-1. Using specific inhibitors, we found that PA stimulation of these tumor cells induced a TLR4-mediated cell invasion. Our results also indicate that the signaling events downstream of TLR4 involved generation of reactive oxygen species, activation of nuclear factor-kappa beta, and secretion and activation of matrix metalloproteinase 9 (MMP-9). Furthermore, PA stimulation decreased the levels of the micro RNA 29c (miR-29c). Of note, while inhibition of miR-29c increased MMP-9 mRNA levels, MMP-9 secretion and activation, and invasiveness, miR-29c mimic abrogated all these PA-stimulated effects. These results strongly suggest that miR-29c could be an attractive potential pharmacological agent for antitumoral therapy in PC.
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Metaloproteinase 9 da Matriz/metabolismo , NF-kappa B/metabolismo , Invasividade Neoplásica , Ácido Palmítico/farmacologia , Neoplasias Pancreáticas/patologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Receptor 4 Toll-Like/metabolismo , Linhagem Celular Tumoral , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias Pancreáticas/enzimologia , Neoplasias Pancreáticas/metabolismoRESUMO
Penile carcinoma (PeCa) is an important public health issue in poor and developing countries, and has only recently been explored in terms of genetic and epigenetic studies. Integrative data analysis is a powerful method for the identification of molecular drivers involved in cancer development and progression. miRNA and mRNA expression profiles followed by integrative analysis were investigated in 23 PeCa and 12 non-neoplastic penile tissues (NPT). Expression levels of eight miRNAs and 10 mRNAs were evaluated in the same set of samples used for microarray and in a validation set of cases (PeCa = 36; NPT = 27). Eighty-one miRNAs and 2,697 mRNAs were identified as differentially expressed in PeCa. Integrative data analysis revealed 255 mRNAs potentially regulated by 68 miRNAs. Using RT-qPCR, eight miRNAs and nine transcripts were confirmed as altered in PeCa. We identified that MMP1, MMP12 and PPARG and hsa-miR-31-5p, hsa-miR-224-5p, and hsa-miR-223-3p were able to distinguish tumors from NPT with high sensitivity and specificity. Higher MMP1 expression was detected as a better predictor of lymph node metastasis than the clinical-pathological data. In addition, PPARG and EGFR were highlighted as potential pathways for targeted therapy in PeCa. The analysis based on HPV positivity (7 of 23 cases) revealed five miRNA and 13 mRNA differentially expressed. Although in a limited number of cases, HPV positive PeCa presented less aggressive phenotype in comparison with negative cases. Overall, an integrative analysis using mRNA and miRNA profiles revealed markers related with tumor development and progression. Furthermore, MMP1 expression level was a predictive marker for lymph node metastasis in patients with PeCa.
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Biomarcadores Tumorais/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias Penianas/genética , RNA Mensageiro/genética , Transdução de Sinais/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Análise por Conglomerados , Diagnóstico Diferencial , Perfilação da Expressão Gênica/métodos , Humanos , Masculino , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 12 da Matriz/genética , Pessoa de Meia-Idade , PPAR gama/genética , Neoplasias Penianas/diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e EspecificidadeRESUMO
In the pathophysiology of spinal cord injury, the secondary biological processes involving changes in gene expression become more important day a day. Within these changes, the expression of different microRNAs has been involved in some of the pathophysiological processes of spinal cord injury. There are several studies that describe the transient expression of microRNA in spinal cord injury, some of them related to inflammation and apoptosis and others to functional recovery and regeneration. MicroRNA may be a potential target for the treatment of spinal cord injury, modifying the processes of inflammation, oxidation, apoptosis, functional recovery and regeneration. It is necessary to continue the study of microRNAs in spinal cord injury, as well as the identification of their target genes and signaling mechanisms involved in its neurological effects. With this, the ultimate goal is the development of effective and safe therapeutic and diagnostic strategies for patients with spinal cord injury.
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MicroRNAs/genética , Medicina Regenerativa/métodos , Traumatismos da Medula Espinal/fisiopatologia , Ambystoma mexicanum , Animais , Apoptose/genética , Regulação da Expressão Gênica , Terapia Genética , Gliose/etiologia , Gliose/genética , Gliose/prevenção & controle , Humanos , Inflamação/genética , Camundongos , Camundongos Transgênicos , MicroRNAs/antagonistas & inibidores , MicroRNAs/biossíntese , Modelos Animais , Regeneração Nervosa/genética , Oligonucleotídeos/uso terapêutico , Oxirredução , Ratos , Recuperação de Função Fisiológica , Medula Espinal/metabolismo , Medula Espinal/fisiologia , Traumatismos da Medula Espinal/epidemiologia , Traumatismos da Medula Espinal/genética , Traumatismos da Medula Espinal/terapiaRESUMO
The functional role of IGFBP5 in breast cancer is complicated. Experimental and bioinformatics studies have shown that IGFBP5 is targeted by miR-140-5p and miR-193b, although this has not yet been proven in clinical samples. The aim of this study was to evaluate the expression of miR-140-5p and miR-193b in breast cancer and adjacent normal tissue and assess its correlation with IGFBP5 and the clinicopathological characteristics of the tumors. IGFBP5 protein expression was analyzed immunohistochemically and IGFBP5, miR-140 and miR-193b mRNA expression levels were analyzed with real-time RT-PCR. Tumor tissue had higher miR-140-5p expression than adjacent normal tissue (p = 0.015). Samples with no immunohistochemical staining for IGFBP5 showed increased miR-140-5p expression (p = 0.009). miR-140-5p expression was elevated in invasive ductal carcinomas (p = 0.002), whereas basal-like tumors had decreased expression of miR-140-5p compared to other tumors (p = 0.008). Lymph node-positive samples showed an approximately 13-fold increase in miR-140-5p expression compared to lymph node-negative tissue (p = 0.049). These findings suggest that miR-140-5p, but not miR-193b, could be an important determinant of IGFBP5 expression and clinical phenotype in breast cancer patients. Further studies are needed to clarify the expressional regulation of IGFBP5 by miR-140-5p.
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Renal ischemia-reperfusion injury (IRI) is a common cause of renal dysfunction and renal failure. Histone/protein deacetylases (HDACs) regulate gene accessibility and higher order protein structures and may alter cellular responses to a variety of stresses. We investigated whether use of pan- and class-specific HDAC inhibitors (HDACi) could improve IRI tolerance in the kidney. Using a model of unilateral renal IRI, we investigated early renal function after IRI, and calculated fibrosis after IRI using an automated scoring system. We found that pan-HDAC inhibition using trichostatin (TSA) yielded significant renal functional benefit at 24-96 hours (p < 0.001). Treated mice developed significantly less fibrosis at 30 days (p < 0.0004). Class I HDAC inhibition with MS-275 yielded similar effects. Protection from fibrosis formation was also noted in a cold ischemia transplant model (p < 0.008) with a trend toward improved cold ischemic survival in TSA-treated mice. These effects were not accompanied by induction of typical ischemic tolerance pathways or by priming of heat shock protein expression. In fact, heat shock protein 70 deletion or overexpression did not alter renal ischemia tolerance. Micro-RNA 21, known to be enhanced in vitro in renal tubular cells that survive stress, was enhanced by treatment with HDACi, pointing to possible mechanism.
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Fibrose/prevenção & controle , Histonas/metabolismo , Isquemia/prevenção & controle , Rim/irrigação sanguínea , Traumatismo por Reperfusão/prevenção & controle , Animais , Feminino , Inibidores de Histona Desacetilases/farmacologia , Camundongos , Camundongos Endogâmicos C57BLRESUMO
The search for biomarkers to characterize prostate cancer aggressiveness has been the objective for the majority of researchers involved with the most prevalent tumor in men. MiRNAs are important for the control of many cellular functions and their deregulation is involved with tumor development and progression. To find miRNAs differentially expressed in prostate cancer and their relation to prognostic factors and biochemical recurrence we studied 53 surgical specimens from men who underwent radical prostatectomy, through a microarray analysis using the microarray platform (GeneChip® miRNA Array - Affymetrix) with more than 46,000 probes and 847 mature human miRNAs and transcripts. We defined different as an expression level greater or less than 1.1 with p<0.05. The validation study using qRT-PCR had confirmed miR21 as overexpressed in tumor that have recurred with a risk of 2.5. Transfection of miR-21 using lipid based assay in DU145 cell line, showed decrease in expression of RECK resulting in increase in expression of MMP9. Invasion assay with Matrigel showed increase in tumor cell invasion after miR-21 transfection. We conclude that miR-21 overexpression is related to increased biochemical recurrence after surgical treatment of prostate cancer. And the negative control of RECK results in overexpression of MMP9 promotes increasing tumor cell invasion supporting miR-21 as an oncomiR related to aggressiveness in prostate cancer.
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OBJECTIVE: MicroRNAs are noncoding RNA molecules involved in the development and progression of tumors. We have found that miRNA-100 is underexpressed in metastatic prostate cancer compared to localized disease. Conversely higher levels of miR-100 are related to biochemical recurrence after surgery. This suggests that miR-100 may be a context-dependent miRNA, acting as oncogene or tumor suppressor miRNA. Our aim is to demonstrate the role of miR-100 in the control of predicted target genes in prostate cancer cell lines. METHODS: Cell lines DU145 and PC3 were transfected with miR-100, antimiR-100 and after 24 h and 48 h of exposure, qRT-PCR and western blot were performed for mTOR, FGFR3, THAP2, SMARCA5 and BAZ2A. RESULTS: There was reduction in mTOR (p = 0.025), THAP2 (p = 0.038), SMARCA5 (p = 0.001) and BAZ2A (p = 0.006) mRNA expression in DU145 cells after exposure to miR-100. In PC3 cells, mTOR expression was decreased by miR-100 (p = 0.01). There was a reduction in the expression levels of proteins encoded by studied genes, ranging from 34% to 69%. CONCLUSIONS: We demonstrate that miR-100 is a context-dependent miRNA controlling BAZ2, mTOR, FGFR3, SMARCA5 and THAP2 that might be involved in PC progression. The elucidation of the roles of miRNAs in tumors is important because they can be used as therapeutic targets in the future. .
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Humanos , Masculino , MicroRNAs/fisiologia , Neoplasias da Próstata/genética , Western Blotting , Linhagem Celular Tumoral , Progressão da Doença , Regulação Neoplásica da Expressão Gênica , Marcação de Genes , MicroRNAs/farmacologia , Valor Preditivo dos Testes , Neoplasias da Próstata/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de TempoRESUMO
OBJECTIVE: The aim of this study is to verify the expression of proteins that are controlled by miR-let7c, 100 and 218 using immunohistochemistry in tissue microarray representative of localized and metastasized the lymph nodes and bone prostate cancer. METHODS: To verify the expression of proteins that are controlled by miR-let7c (C-MYC, BUB1, RAS) 100 (SMARCA5, RB) and 218 (LAMB3) and cell proliferation (Ki-67) we used immunohistochemistry and computerized image system ImageJ MacBiophotonics in three tissue microarrays representative of localized prostate cancer and lymph node and bone metastases. miRNA expression was evaluated by qRT-PCR using 60 paraffin blocks to construct the tissue microarray representative of localized disease. RESULTS: RAS expression was increased in localized prostate cancer and bone metastases compared to the lymph nodes (p=0.017). RB showed an increase in expression from localized prostate cancer to lymph node and bone metastasis (p=0.036). LAMB3 was highly expressed in localized and lymph node metastases (p<0.001). Cell proliferation evaluated by Ki-67 showed an increase from localized prostate cancer to metastases (p<0.001). We did not found any relationship between C-MYC (p=0.253), BUB1 (p=0.649) and SMARCA5 (p=0.315) protein expression with prognosis or tumor behavior. CONCLUSION: We found that the expression of RAS, RB, LAMB3 and Ki-67 changed in the different stages of prostate cancer. Furthermore, we confirmed the overexpression of the miRNAs let7c, 100 and 218 in localized prostate cancer but failed to show the control of protein expression by the putative controller miRNAs using immunohistochemistry. .
Assuntos
Adulto , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Ósseas/secundário , MicroRNAs/metabolismo , Proteínas de Neoplasias/fisiologia , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Adenosina Trifosfatases/metabolismo , Moléculas de Adesão Celular/metabolismo , Proteínas Cromossômicas não Histona/metabolismo , Regulação Neoplásica da Expressão Gênica , Imuno-Histoquímica , /metabolismo , Metástase Linfática , MicroRNAs/genética , MicroRNAs/fisiologia , Proteínas de Neoplasias/metabolismo , Prognóstico , Neoplasias da Próstata/genética , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Proto-Oncogênicas c-myc/metabolismo , /metabolismo , Proteína do Retinoblastoma/metabolismoRESUMO
INTRODUCTION: Models of the multistep process related to cancer progression have been designed for many cancers including prostate. The aim of this study is to propose a new model including a possible role for recently described micro RNAs in prostate cancer (CaP) progression. METHODS: Sixty-three patients underwent radical prostatectomy to treat localized prostate carcinoma. The specimens of 15 patients were representative of high grade prostate intraepithelial neoplasia (HGPIN). Fourteen specimens represented localized favorable CaP, and 34 unfavorable, mostly non-organ-confined disease. Representing the advanced disease we studied 4 metastatic androgen-independent CaP and 2 cell lines. Micro RNAs were isolated using the mirVana miRNA Isolation kit and cDNA was obtained using the TaqMan miRNA Reverse Transcription kit to the miRNAs: hsa-miR-let7c, hsa-miR-15a, hsa-miR-16, hsa-miR-21, hsa-miR-25, hsa-miR-32, hsa-miR-100, hsa-miR-143, hsa-miR-145, hsa-miR-146a, hsa-miR-191, hsa-miR-199a, hsa-miR-206, and hsa-miR-218. Quantitative RT-PCR was carried out using the ABI 7500 Fast Real-Time PCR System and the TaqMan Universal PCR Master Mix. miRNA expression levels were measured by relative quantification, and fold expression changes were determined by the 2(-ΔΔCT) method. The small nucleolar RNA RNU43 was used as an endogenous control. RESULTS: Except for miR-21 and miR-206, the expression levels of all miRNAs significantly changed during the progression of CaP. Interestingly, there was a significant global loss of miRNA expression between HGPIN and metastasis at 2 important steps. The first was related to the transition from HGPIN to invasive adenocarcinoma, and the second was related to the transition from localized to metastatic adenocarcinomas. CONCLUSION: Through the analysis of 14 miRNAs in 4 groups of prostate lesions, which reproduced the progression of CaP, we showed that there is a global loss of miRNA expression at 2 distinct steps. The first related to the transition between HGPIN and localized invasive carcinoma, and the second associated with the transition from localized to metastatic CaP. The importance of our study is in the identification of possible miRNAs and miRNA-targeted genes involved in the progression of prostate carcinogenesis that may help the development of potential diagnostic or prognostic markers as well as the design of new target therapies.