Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 100.245
Filtrar
1.
Talanta ; 237: 122927, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34736664

RESUMO

Abnormal expression of microRNAs is greatly associated with the occurrence of various cancer types, revealing great potential of microRNA as biomarkers for cancer diagnosis and prognosis. Herein, a MXene-MoS2 heterostructure enhancing electrochemical biosensor coupled with catalytic hairpin assembly (CHA) amplification approach for label-free determination of microRNA-21 (miR-21) was successfully assembled. In particular, the unique micro-nano heterostructure with large specific area and favorable electroconductivity exhibited the ability of excellent confinement effect. Thus, rendered the MXene-MoS2 heterostructure the ability to trigger more target recycling reaction, giving new vitality to the traditional CHA amplification method. Meanwhile, thionine (Thi) and gold nanoparticles (AuNPs) were anchoring at the surface of MXene-MoS2 heterostructure, respectively, empowered the sensor the capability of capture probes fixation and miR-21 label-free determination. When numerous electronegative double-stranded DNA generated, the electron transfer was greatly hindered, resulting in signal decrease. Accordingly, the design denoted a broad dynamic range from 100 fM to 100 nM and a detection limit of about 26 fM, comparable or lower than previous reported methods for miR-21 detection. Furthermore, the sensing platform supplied satisfactory selectivity, reproducibility and stability towards the miR-21 detection. The real sample determination also showed a promising performance under clinical circumstance. Finally, from the clinical standpoint, the proposed biosensor is a considerable platform toward early disease detection and monitoring.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , MicroRNAs , Catálise , Técnicas Eletroquímicas , Ouro , Limite de Detecção , Molibdênio , Reprodutibilidade dos Testes
2.
Talanta ; 237: 122969, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34736693

RESUMO

In this work, a novel electrochemiluminescence (ECL) sensor has been developed to detect miRNA-210 in the serum of triple negative breast cancer (TNBC) patients. The luminous MoS2 nanosheets were synthesized via the solvothermal method and served as ECL emitters for the first time. As a result, the ECL properties of as-prepared MoS2 nanosheets were significantly improved. Furthermore, the biomimetic magnetic vesicles were used as capture platform in the ECL sensing strategy. Due to the highly efficient fluidity and magnetic property, the biomimetic vesicles with hairpin aptamers can capture target gene in the serum. After magnetic separation, the captured miRNA-210 can trigger the target-catalyzed hairpin assembly (CHA) sensing process on the magnetic electrode and hybridize MoS2 nanosheets labeled probe DNA. The concentration of miRNA-210 can be quantified by the ECL enhancement of the MoS2 nanosheets. This approach has achieved the sensitive detection for miRNA-210 in a range from 1 fM to 100 pM with the detection limit of 0.3 fM. The luminous MoS2 nanosheets-based ECL sensing system with the biomimetic vesicles would provide a new pathway to explore 2D nanomaterials for developing a wide range of bioanalytical applications.


Assuntos
Técnicas Biossensoriais , MicroRNAs , Biomimética , Técnicas Eletroquímicas , Humanos , Limite de Detecção , Medições Luminescentes , Molibdênio
3.
Anal Chim Acta ; 1189: 339182, 2022 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-34815041

RESUMO

Dysregulation of MicroRNAs (miRNAs) cause various diseases in humans, and developing reliable methods to detect miRNAs is critical for molecular diagnostics and personalized medicine. This study developed a toehold-mediated target invasion combined with duplex-specificity nuclease (DSN)-assisted cyclic signal amplification fluorescent sensor. Herein, we take advantage of toehold-mediated target invasion process to ensure the high selectivity of miRNA determination, coupled with the unique cleavage properties of DSN to improve the sensitivity of the strategy significantly. Throughout the assay, the whole procedure of detection the target let-7a has a limit of detection (LOD) as low as 9.00 fM and an excellent linear range from 1 pM to 100 nM for no more than 60 min. The assay shows reasonable specificity in detecting mismatched miRNAs and can realize single-base discrimination in the let-7 families. Finally, the developed method was applied to detect the miRNAs extracted from human serum. The results were consistent with those based on the quantitative reverse transcription-polymerase chain reaction(qRT-PCR) method, which shows great potential application value in clinical molecular diagnostics and biological research.


Assuntos
Técnicas Biossensoriais , MicroRNAs , Endonucleases , Humanos , Limite de Detecção , MicroRNAs/genética , Técnicas de Amplificação de Ácido Nucleico
4.
Anal Chim Acta ; 1189: 338701, 2022 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-34815048

RESUMO

Highly sensitive and specific imaging of low-level microRNAs (miRNAs) in cytoplasm is vital for early diagnosis of cancers. In this work, we have developed the amplification strategies for miRNA-155 detection based on the combination the nicked rolling circle amplification (N-RCA) and catalyzed hairpin assembly (CHA). In this system, the target miRNA-155 acts as a polymerase primer to activate N-RCA to produce nicked fragment1 (NF1) and NF2. NF1 acted as new primer could further initiate a new N-RCA reaction over and over. Then, the NF2s could serve as triggers to induce the CHA reaction, and the Y-shaped DNA nanostructure (Y-SDN) was formed. Thus, an amplified fluorescence signal was obtained based on the multiple amplification. Under the optimized experimental conditions, a high sensitivity with a detection limit as low as 1.8 pM at 3σ miRNA-155 and excellent specificity in buffer condition have been achieved by applying this method. Meanwhile, the proposed method enables the application in miRNA-155 detection in human serum. Moreover, we have shown that the method performs well for the intracellular miRNA-155 imaging in cellular environments. Therefore, the present strategy was expected to apply into the clinical disease diagnosis effectively.


Assuntos
Técnicas Biossensoriais , MicroRNAs , Nanoestruturas , DNA , Testes Diagnósticos de Rotina , Fluorescência , Humanos , Limite de Detecção , Técnicas de Amplificação de Ácido Nucleico
5.
Environ Pollut ; 292(Pt A): 118332, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34637826

RESUMO

With the continued increase of global ammonia emission, the damage to human or animal caused by ammonia pollution has attracted wide attention. The noncoding RNAs have been reported to regulate a variety of biological processes under different environmental stimulation via ceRNA (competing endogenous RNA) networks. Autophagy is a hallmark of tissue damage from air pollution. However, the specific role of circular RNAs (circRNAs) in the injury of intestinal tissue caused by autophagy remains unclear. Here, we established 42-days old ammonia-exposed broiler models and observed that autophagy flux in broiler jejunum was activated under ammonia exposure. Meanwhile, a total of eight significantly dysregulated expressed circRNAs were obtained and a circRNAs-miRNAs-genes interaction networks were constructed by bioinformatics analysis. Furthermore, an axis named circRNA-IGLL1/miR-15a/RNF43 was predicted to participate in the excessive autophagy by targeting RNF43. The target relationship was proved by dual-luciferase reporter assay in vitro. Mechanistically, downregulated circRNA-IGLL1 could suppress the expression of RNF43 in ammonia-exposed jejunum and the Wnt/ß-catenin pathway was activated. Inhibition of miR-15a reversed autophagy caused by downregulated circRNA-IGLL1. CircRNA-IGLL1 could competitively bind miR-15a to regulate RNF43 expression, thus modulating the occurrence of autophagy. Taken together, our results showed that circRNA-IGLL1/miR-15a/RNF43 axis is involved in ammonia-induced intestinal autophagy in broilers.


Assuntos
MicroRNAs , RNA Circular , Amônia/toxicidade , Animais , Autofagia , Galinhas , Humanos , Jejuno , MicroRNAs/genética , Ubiquitina-Proteína Ligases , beta Catenina
6.
Environ Pollut ; 292(Pt B): 118387, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34673158

RESUMO

Ambient particulate air pollution is a risk factor for cardiovascular and respiratory disease, yet the biological mechanisms underlying this association are not well understood. The current study aimed to investigate the mediation role of microRNAs on the association between personal PM2.5 exposure and blood pressure and lung function. One hundred and twenty adults (60 truck drivers and 60 office workers) aged 18-46 years were assessed on the June 15, 2008 and at follow-up (1- to 2-weeks later). MicroRNAs were extracted from the peripheral blood samples. Compared to truck drivers, there is a significant increase in FEF25-75, FEV1, and FEV1/FVC and a decrease in PM2.5 in office workers (all p < 0.05). According to the Bonferroni corrected threshold p-value < 6.81 × 10-5 (0.05/734) used, personal PM2.5 data showed a significant positive association with miR-644 after the adjustment for age, BMI, smoking status, and habitual alcohol use. The mediation effect of miR-644 on the association between personal PM2.5 exposure and FEF25-75 [B (95%CI) = -1.342 (-2.810, -0.113)], PEF [B (95%CI) = -1.793 (-3.926, -0.195)], and FEV1/FVC [B (95%CI) = -0.119‰ (-0.224‰, -0.026‰)] was significant only for truck drivers after the adjustment for covariates. There were no similar associations with blood pressure. These results demonstrate microRNAs to potentially mediate association of PM2.5 with lung function. Subsequent studies are needed to further elucidate the potential mechanisms of action by which the mediation effect of microRNAs is achieved with this process.


Assuntos
Poluentes Atmosféricos , Poluição do Ar , MicroRNAs , Adulto , Poluentes Atmosféricos/análise , Poluição do Ar/efeitos adversos , Poluição do Ar/análise , Pressão Sanguínea , Exposição Ambiental/análise , Humanos , Pulmão , MicroRNAs/genética , Material Particulado/análise
7.
Talanta ; 236: 122846, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635236

RESUMO

Simultaneous detection of multiple microRNAs (miRNAs) with high sensitivity can give accurate and reliable information for clinical applications. By uniformly anchoring hairpin probes on the surface of DNA nanolantern, a three-dimensional DNA nanostructure contains abundant and adjustable modification sites, highly integrated DNA nanoprobes were designed and developed as catalytic hairpin assembly (CHA)-based signal amplifiers for enzyme-free signal amplification detection of target miRNAs. The nanolantern-based CHA (NLC) amplifiers, which were facilely prepared via a simple "one-pot" annealing method, showed enhanced biostability, improved cell internalization efficiency, accelerated CHA reaction kinetics, and increased signal amplification capability compared to the single-stranded DNA hairpin probes used in traditional CHA reaction. By co-assembling multiple hairpin probes on a DNA nanolantern surface, as-prepared NLC amplifiers were demonstrated to work well for highly sensitive and specific imaging, expression level fluctuation analysis of two miRNAs in living cells, and miRNAs-guided tumor imaging in living mice. The proposed DNA nanolantern-based nanoamplifier strategy might provide a feasible way to promote the cellular and in vivo applications of nucleic acid probes.


Assuntos
Técnicas Biossensoriais , MicroRNAs , Animais , Catálise , DNA/genética , Camundongos , MicroRNAs/genética , Sondas de Ácido Nucleico
8.
J Hazard Mater ; 421: 126760, 2022 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-34396970

RESUMO

Large amounts of epidemiological evidence have confirmed the atmospheric particulate matter (PM2.5) exposure was positively correlated with the morbidity and mortality of respiratory diseases. Nevertheless, its pathogenesis remains incompletely understood, probably resulting from the activation of oxidative stress, inflammation, altered genetic and epigenetic modifications in the lung upon PM2.5 exposure. Currently, biomarker investigations have been widely used in epidemiological and toxicological studies, which may help in understanding the biologic mechanisms underlying PM2.5-elicited adverse health outcomes. Here, the emerging biomarkers to indicate PM2.5-respiratory system interactions were summarized, primarily related to oxidative stress (ROS, MDA, GSH, etc.), inflammation (Interleukins, FENO, CC16, etc.), DNA damage (8-OHdG, γH2AX, OGG1) and also epigenetic modulation (DNA methylation, histone modification, microRNAs). The identified biomarkers shed light on PM2.5-elicited inflammation, fibrogenesis and carcinogenesis, thus may favor more precise interventions in public health. It is worth noting that some inconsistent findings may possibly relate to the inter-study differentials in the airborne PM2.5 sample, exposure mode and targeted subjects, as well as methodological issues. Further research, particularly by -omics technique to identify novel, specific biomarkers, is warranted to illuminate the causal relationship between PM2.5 pollution and deleterious lung outcomes.


Assuntos
Poluentes Atmosféricos , MicroRNAs , Poluentes Atmosféricos/análise , Poluentes Atmosféricos/toxicidade , Biomarcadores , Humanos , Pulmão , Estresse Oxidativo , Material Particulado/análise , Material Particulado/toxicidade
9.
Chemosphere ; 286(Pt 1): 131614, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34325257

RESUMO

Particulate matter (PM)-induced airway inflammation contributes to the development and exacerbation of chronic airway diseases. Circular RNA (circRNA) is a new class of non-coding RNA that participates in gene regulation in various respiratory diseases, but the regulatory role of circRNA in PM-induced airway inflammation has not been fully elucidated. In this study, we performed the human circRNA microarray to reveal differentially expressed circRNAs in PM-induced human bronchial epithelial cells (HBECs). A total of 176 upregulated and 15 downregulated circRNAs were identified. Of these, a new circRNA termed circTXNRD1 was upregulated by PM exposure in a dose- and time-dependent manner. Knockdown of circTXNRD1 significantly attenuated PM-induced expression of proinflammatory cytokine interleukin 6 (IL-6). CircRNA pull-down, dual-luciferase reporter assay and fluorescence in situ hybridization showed that circTXNRD1 acted as an endogenous sponge to decrease miR-892a levels in HBECs. Downregulation of miR-892a could increase cyclooxygenase-2 (COX-2) expression and eventually promote IL-6 secretion in PM-induced HBECs. Taken together, our findings reveal circTXNRD1 as a novel inflammatory mediator in PM-induced inflammation in HBECs via regulating miR-892a/COX-2 axis. These results provide new insight into the biological mechanism underlying PM-induced inflammation in chronic airway diseases.


Assuntos
MicroRNAs , RNA Circular , Ciclo-Oxigenase 2/genética , Células Epiteliais , Humanos , Hibridização in Situ Fluorescente , Inflamação/induzido quimicamente , Inflamação/genética , MicroRNAs/genética , Material Particulado/toxicidade , RNA/genética
10.
Food Chem ; 368: 130816, 2022 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-34416489

RESUMO

Acrylamide (AA), a potential carcinogen, is commonly formed in foods rich in carbohydrates at high heat. It is known that AA-induced mitochondrial dysfunction is responsible for its toxicity. Previously we found AA exposure increased miR-27a-5p expression in livers of SD rats. Here, the regulation mechanism of miR-27a-5p in mitochondrial dysfunction was investigated in rat liver cell lines (IAR20) and SD rats. The results showed that the overexpressed miR-27a-5p contributes to modulating mitochondrial dysfunction and Btf3 is identified as its target gene. The knockdown of Btf3 increases the cleaved PARP1 level and the phosphorylation of ATM and p53, which results in mitochondria-dependent apoptosis. Therefore, the miR-27a-5p-Btf3-ATM-p53 axis might play a vital role in the promotion of AA-induced cell apoptosis through disrupting mitochondrial structure and function. This would provide a potential target for the assessment and intervention of AA toxicity.


Assuntos
MicroRNAs , Acrilamida/toxicidade , Animais , Apoptose , MicroRNAs/genética , Mitocôndrias/genética , Ratos , Ratos Sprague-Dawley
11.
Environ Pollut ; 292(Pt B): 118452, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34737026

RESUMO

The present study aims to determine the associations of multiple plasma metal levels and plasma microRNAs (miRNAs) with diabetes risk, and further explore the mediating effects of plasma miRNAs on the associations of plasma metal with diabetes risk. We detected plasma levels of 23 metals by inductively coupled plasma mass spectrometry (ICP-MS) among 94 newly diagnosed and untreated diabetic cases and 94 healthy controls. The plasma miRNAs were examined by microRNA Array screening and Taqman real-time PCR validation among the same study population. The multivariate logistic regression models were employed to explore the associations of plasma metal and miRNAs levels with diabetes risk. Generalized linear regression models were utilized to investigate the relationships between plasma metal and plasma miRNAs, and mediation analysis was used to assess the mediating effects of plasma miRNAs on the relationships between plasma metals and diabetes risk. Plasma aluminum (Al), titanium (Ti), copper (Cu), zinc (Zn), selenium (Se), rubidium (Rb), strontium (Sr), barium (Ba), and Thallium (Tl) levels were correlated with elevated diabetic risk while molybdenum (Mo) with decreased diabetic risk (P < 0.05 after FDR multiple correction). MiR-122-5p and miR-3141 were positively associated with diabetes risk (all P < 0.05). Ti, Cu, and Zn were positively correlated with miR-122-5p (P = 0.001, 0.028 and 0.004 respectively). Ti, Cu, and Se were positively correlated with miR-3141 (P = 0.003, 0.015, and 0.031 respectively). In addition, Zn was positively correlated with miR-193b-3p (P = 0.002). Ti was negatively correlated with miR-26b-3p (P = 0.016), while Mo and miR-26b-3p were positively correlated (P = 0.042). In the mediation analysis, miR-122-5p mediated 48.0% of the association between Ti and diabetes risk. The biological mechanisms of the association are needed to be explored in further studies.


Assuntos
Diabetes Mellitus Tipo 2 , MicroRNAs , Cobre , Humanos , Metais , Titânio
12.
Theriogenology ; 177: 84-93, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34687940

RESUMO

MicroRNAs (miRNAs) are ∼22 nt RNAs that direct post-transcriptional repression of mRNA targets in diverse eukaryotic lineages. Granulosa cells (GCs) are the earliest differentiated follicular somatic cells. From the initiation of primordial follicles, their differentiation and growth are closely related to the development of follicles. The research on follicular development mostly focused on the granular layer, as well as the hormone synthesis induced by granulosa cell differentiation before and after follicular selection. In this study, we evaluated the effects of miR-23b-3p on chicken granulosa cells, including granulosa cell proliferation and steroid hormone synthesis. Elevated expression of miR-23b-3p significantly inhibited granulosa cell proliferation and steroid hormone synthesis, but did not affect apoptosis. Furthermore, it was observed that the forecast growth differentiation factor 9 (GDF9) is a target gene of miR-23b-3p and miR-23b-3p can down-regulate expression of GDF9. Overall, this study demonstrated that miR-23b-3p can regulate the proliferation and steroid hormone synthesis of chicken granulosa cells by inhibiting the expression of GDF9.


Assuntos
Fator 9 de Diferenciação de Crescimento , MicroRNAs , Animais , Proliferação de Células , Galinhas/genética , Feminino , Fator 9 de Diferenciação de Crescimento/genética , Hormônios , MicroRNAs/genética , Esteroides
13.
BMC cancer ; 21(1): 575-678, May., 2021. ilus, graf, tab
Artigo em Inglês | Sec. Est. Saúde SP, CONASS, SESSP-IDPCPROD, Sec. Est. Saúde SP | ID: biblio-1224518

RESUMO

BACKGROUND: No biomarker is available for identifying cancer patients at risk of developing nephrotoxicity when treated with cisplatin. METHODS: We performed microRNA (miRNA) sequencing using plasma collected 5 days after cisplatin treatment (D5) from twelve patients with head and neck cancer with and without nephrotoxicity (grade ≥ 2 increased serum creatinine). The most differentially expressed miRNAs between the two groups were selected for quantification at baseline and D5 in a larger cohort of patients. The association between miRNAs and nephrotoxicity was evaluated by calculating the odds ratio (OR) from univariate logistic regression. Receiver operating characteristic curves (ROC) were used to estimate the area under the curve (AUC), sensitivity, and specificity. RESULTS: MiR-3168 (p = 1.98 × 10− 8 ), miR-4718 (p = 4.24 × 10− 5 ), and miR-6125 (p = 6.60 × 10− 5 ) were the most differentially expressed miRNAs and were further quantified in 43, 48, and 53 patients, respectively. The baseline expression of miR-3168 (p = 0.0456, OR = 1.03, 95% CI: 1.00­1.06) and miR-4718 (p = 0.0388, OR = 1.56, 95% CI: 1.03­ 2.46) were associated with an increased risk of nephrotoxicity, whereas miR-6125 showed a trend (p = 0.0618, OR = 1.73, 95% CI: 0.98­3.29). MiR-4718 showed the highest AUC (0.77, 95% CI: 0.61­0.93) with sensitivity of 66.76 and specificity of 79.49. CONCLUSIONS: We have provided evidence of baseline plasmatic expression of miR-3168, miR-6125, and miR-4718 as potential predictors of cisplatin-induced nephrotoxicity.


Assuntos
Cisplatino , MicroRNAs , Nefropatias , Neoplasias
14.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 43(5): 669-676, 2021 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-34728027

RESUMO

Objective To explore the effect of miR-145-5p on the proliferation and apoptosis of human ovarian cancer cells and the possible molecular mechanisms involved.Methods Real-time quantitative PCR was performed to detect the expression of miR-145-5p in ovarian epithelial cells and ovarian cancer cells.CCK-8 and flow cytometry were used to detect the effects of miR-145-5p overexpression on the proliferation and apoptosis of ovarian cancer cells.TargetScan was employed to predict the target genes of miR-145-5p.Western blotting,dual luciferase reporter assay and rescue experiment were employed to predict and verify the underlying molecular mechanism of miR-145-5p function.Results The expression of miR-145-5p in ovarian cancer cells was significantly lower than that in normal ovarian epithelial cells(t=4.345,P=0.049).Compared with the control group,the overexpression of miR-145-5p reduced the proliferation rate(t=-15.790,P<0.001)and increased the apoptosis rate(t=5.433,P=0.032)of ovarian cancer cells.ARK5 was predicted as the direct target gene of miR-145-5p(t=4.583,P=0.010).The cells with ARK5 overexpression showed increased proliferation rate(t=27.290,P<0.001)and decreased apoptosis rate(t=-8.241,P=0.001).The overexpression of miR-145-5p can down-regulate the mRNA(t=-12.824,P<0.001)and protein(t=-4.792,P=0.001)levels of ARK5.The rescuing expression of ARK5 significantly offset the inhibitory effects of miR-145-5p on cell proliferation(t=15.580,P=0.004)and apoptosis(t=-12.470,P=0.006).Conclusion miR-145-5p may inhibit the proliferation and promote the apoptosis of ovarian cancer cells by targeting ARK5.


Assuntos
MicroRNAs , Neoplasias Ovarianas , Apoptose/genética , Carcinoma Epitelial do Ovário/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , MicroRNAs/genética , Neoplasias Ovarianas/genética
15.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 43(5): 815-821, 2021 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-34728045

RESUMO

In recent years,microRNAs(miRNAs)have been detected at different stages of follicular development and in different cells of follicles.Extracellular vesicle(EV)-derived miRNAs have also been detected in the follicular fluid of mature follicles.miRNAs participate in the regulation of normal follicular development,and the regulation disorder may lead to the occurrence of some ovarian diseases.In order to further systematically elucidate the regulatory mechanism of miRNAs on follicular development and find suitable EV-derived miRNAs that can predict oocyte development,we reviewed the functions of miRNAs in follicular development from the perspectives of granulosa cell development,oocyte development,and hormone synthesis.


Assuntos
MicroRNAs , Feminino , Líquido Folicular , Células da Granulosa , Humanos , MicroRNAs/genética , Oogênese , Folículo Ovariano
16.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 38(11): 1091-1096, 2021 Nov 10.
Artigo em Chinês | MEDLINE | ID: mdl-34729750

RESUMO

OBJECTIVE: To explore the effect of HNF1A-AS1 on the proliferation, migration and invasion of IL-6-induced hemangioendothelial cells (HemEC) and possible mechanism. METHODS: RT-qPCR was used to detect the expression level of HNF1A-AS1 and miR-363-3p in the tumor tissue and adjacent normal skin tissue from 35 patients with hemangioma. Pearson correlation was used to analyze the correlation between the expression of HNF1A-AS1 and miR-363-3p in tumor tissues. HemEC were isolated and cultured in vitro.Dual luciferase reporter gene experiment was used to study the regulatory effect between HNF1A-AS1 and miR-363-3p. IL-6 was added to HemEC transfected with si-NC, si-HNF1A-AS1, si-HNF1A-AS1 and anti-miR-NC, or si-HNF1A-AS1 and anti-miR-363-3p, respectively. CCK-8 method and clone formation experiment were used to detect cell proliferation in each group. Transwell method was used to detect cell migration and invasion in each group. Western blotting was used to detect the expression of Ki67, MMP-2 and MMP-9 proteins in each group. RESULTS: Compared with normal skin tissues, the expression of IL-6 mRNA in hemangioma tissues was increased (P<0.05), and the expression of IL-6 mRNA in the proliferative phase was lower than that in the degenerative phase (P<0.05). Expression of HNF1A-AS1 in hemangioma tissue was increased (P<0.05), while that of miR-363-3p was decreased (P<0.05), and the two were negatively correlated (r=-0.758, P<0.05). HNF1A-AS1 down-regulated the expression of miR-363-3p in HemEC.IL-6 promoted the expression of HNF1A-AS1, OD value, number of colonies, number of migration and invasion of HemEC cells, and the expression of Ki67, MMP-2 and MMP-9proteins (P<0.05), while reduced the expression of miR-363-3p (P<0.05). Down-regulating si-HNF1A-AS1 reduced the IL-6-induced HemEC cell OD value, colony numbers, migration and invasion and the expression of Ki67, MMP-2 and MMP-9 proteins (P<0.05). Down-regulating miR-363-3p attenuated the inhibitory effect of down-regulating si-HNF1A-AS1 on the proliferation, migration and invasion of HemEC cells induced by IL-6 (P<0.05). CONCLUSION: Expression of HNF1A-AS1 is increased in hemangioma tissues. Down-regulating HNF1A-AS1 may inhibit proliferation, migration and invasion of IL-6-induced hemangioma endothelial cells by targeted up-regulation of miR-363-3p.


Assuntos
Hemangioma , MicroRNAs , RNA Longo não Codificante , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Células Endoteliais , Regulação Neoplásica da Expressão Gênica , Hemangioma/genética , Fator 1-alfa Nuclear de Hepatócito/genética , Humanos , Interleucina-6/genética , MicroRNAs/genética
17.
Acta Chir Orthop Traumatol Cech ; 88(5): 344-353, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34738893

RESUMO

PURPOSE OF THE STUDY The aim of the study was to determine miR-146a-5p, miR-223-3p and miR-23a-3p by an enzyme immunoassay in patients with inflammatory and non-inflammatory joint effusion and to verify the usefulness of these miRNAs as biomarkers of joint inflammation. MATERIAL AND METHODS Synovial fluid (SF) samples were collected from 82 patients. The group consisted of 60 non-inflammatory, 11 inflammatory-non-pyogenic, 11 inflammatory-pyogenic SF. SF miRNA was isolated by RNA Isolation Kit Plasma/Serum. The concentrations of miRNA were determined by enzyme-linked immunosorbent assays (ELISA), C-reactive protein, interleukin-6 and procalcitonin on automatic analyser, presepsin on POCT system, interleukin-1 and human neutrofil defensins 1-3 by ELISA. RESULTS A statistically significant negative correlation was found between miR-146-5p and miR-223-3p, WBC, IL-1ß, IL-6 and CRP (P < 0.05) in all groups; a statistically significant positive correlation was found between miR-223-3p and miR-23a-3p, WBC, PMN, IL-1beta, IL-6 and HNP1-3, as well as a positive correlation of miR-23a-3p with IL-1ß, IL-6 and HNP1-3. A statistically significant difference was found between miR-146a-5p, miR-223-3p and miR-23a-3p and individual SF groups, P = 0.006, P < 0.001, respectively. PMN, WBC, Il-1ß, IL-6, HNP 1-3 predicted the inflammatory processes with excellent diagnostic power (AUC > 0.9). The clinical relevance expressed by effect size was the strongest in miR-223-3p, PMN, IL-1 , HNP 1-3 between non-inflammatory and inflammatory-pyogenic group. CONCLUSIONS Our study quantified the SF miRNA by ELISA. We have shown that miR-146a-5p, miR-223-3p and miR-23a-3p can be an important group of biomarkers for the detection and monitoring of various pathophysiological conditions in synovial fluid, including inflammatory conditions. Key words: miRNA, synovial fluid, inflammatory joint disease, enzyme-linked immunosorbent assay.


Assuntos
MicroRNAs , Biomarcadores , Humanos , Receptores de Lipopolissacarídeos , MicroRNAs/genética , Fragmentos de Peptídeos
18.
Planta ; 254(6): 113, 2021 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-34739601

RESUMO

MAIN CONCLUSION: High-throughput sequencing and yeast one and two-hybrid library screening reveal that DKGA2ox1 and miR171f_3 are involved in the regulation of scion dwarfing with 'Nan-tong-xiao-fang-shi' as interstocks. Diospyros kaki Thunb. cv. Nan-tong-xiao-fang-shi ('Nan-tong-xiao-fang-shi') interstocks play a critical role in the scion dwarfing. However, the understanding of the molecular signaling pathways that regulate the scion dwarfing with 'Nan-tong-xiao-fang-shi' as interstocks remain unclear. In this work, the regulatory network in the scion dwarfing with 'Nan-tong-xiao-fang-shi' as interstocks was identified. Using a yeast one-hybrid library screening, luciferase activity analysis, luciferase complementation imaging assays and GFP signal detection, a SPL transcription factor named Diospyros kaki SPL (DKSPL), potentially functioning as a transcriptional activator of the Diospyros kaki GA2ox1 (DKGA2ox1) gene, was identified as a key stimulating factor in the persimmon growth and development. The DKSPL was found in the nucleus, and might play a role in the transcriptional regulation system. A microRNA named miR171f_3 was identified, which might act as a negative regulator of Diospyros kaki SCR (DKSCR) in persimmon. The interactions between DKSCR and seven proteins were experimentally validated with a yeast two-hybrid library screening. Compared to the non-grafted wildtype persimmon, the tissue section of graft union healed well due to the increased expression of cinnamyl-alcohol dehydrogenase. These results indicate that DKGA2ox1 and miR171f_3 may co-promote the scion dwarfing by plant hormone signal transduction pathways.


Assuntos
Diospyros , MicroRNAs , Diospyros/genética , Frutas , MicroRNAs/genética , Reguladores de Crescimento de Plantas , Fatores de Transcrição
19.
BMC Genomics ; 22(Suppl 3): 793, 2021 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-34736408

RESUMO

BACKGROUND: Winter wheat requires prolonged exposure to low temperature to initiate flowering (vernalization). Shoot apical meristem of the crown is the site of cold perception, which produces leaf primordia during vegetative growth before developing into floral primordia at the initiation of the reproductive phase. Although many essential genes for winter wheat cold acclimation and floral initiation have been revealed, the importance of microRNA (miRNA) meditated post-transcriptional regulation in crowns is not well understood. To understand the potential roles of miRNAs in crown tissues, we performed a temporal expression study of miRNAs in crown tissues at the three-leaf stage, winter dormancy stage, spring green-up stage, and jointing stage of winter wheat grown under natural growth conditions. RESULTS: In total, 348 miRNAs belonging to 298 miRNA families, were identified in wheat crown tissues. Among them, 92 differentially expressed miRNAs (DEMs) were found to be significantly regulated from the three-leaf stage to the jointing stage. Most of these DEMs were highly expressed at the three-leaf stage and winter dormancy stage, and then declined in later stages. Six DEMs, including miR156a-5p were markedly induced during the winter dormancy stage. Eleven DEMs, including miR159a.1, miR390a-5p, miR393-5p, miR160a-5p, and miR1436, were highly expressed at the green-up stage. Twelve DEMs, such as miR172a-5p, miR394a, miR319b-3p, and miR9676-5p were highly induced at the jointing stage. Moreover, 14 novel target genes of nine wheat or Pooideae-specific miRNAs were verified using RLM-5' RACE assay. Notably, six mTERFs and two Rf1 genes, which are associated with mitochondrial gene expression, were confirmed as targets of three wheat-specific miRNAs. CONCLUSIONS: The present study not only confirmed the known miRNAs associated with phase transition and floral development, but also identified a number of wheat or Pooideae-specific miRNAs critical for winter wheat cold acclimation and floral development. Most importantly, this study provided experimental evidence that miRNA could regulate mitochondrial gene expression by targeting mTERF and Rf1 genes. Our study provides valuable information for further exploration of the mechanism of miRNA mediated post-transcriptional regulation during winter wheat vernalization and inflorescent initiation.


Assuntos
MicroRNAs , Triticum , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Humanos , Meristema , MicroRNAs/genética , Triticum/genética
20.
J Int Med Res ; 49(11): 3000605211055059, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34772311

RESUMO

OBJECTIVE: To investigate the effect of liraglutide on the browning of white fat and the suppression of obesity via regulating microRNA (miR)-27b in vivo and in vitro. METHODS: Sprague-Dawley rats were fed a high-fat (HF) diet and 3T3-L1 pre-adipocytes were differentiated into mature white adipocytes. Rats and mature adipocytes were then treated with different doses of liraglutide. The mRNA and protein levels of browning-associated proteins, including uncoupling protein 1 (UCP1), PR domain containing 16 (PRDM16), CCAAT enhancer binding protein ß (CEBPß), cell death-inducing DFFA-like effector A (CIDEA) and peroxisome proliferator-activated receptor-γ-coactivator 1α (PGC-1α), were detected using quantitative real-time polymerase chain reaction and Western blotting. RESULTS: Liraglutide decreased body weight and reduced the levels of blood glucose, triglyceride and low-density lipoprotein cholesterol in HF diet-fed rats. Liraglutide increased the levels of UCP1, PRDM16, CEBPß, CIDEA and PGC-1α in vivo and vitro. The levels of miR-27b were upregulated in HF diet-fed rats, whereas liraglutide reduced the levels of miR-27b. In vitro, overexpression of miR-27b decreased the mRNA and protein levels of UCP1, PRDM16, CEBPß, CIDEA and PGC-1α. Transfection with the miR-27b mimics attenuated the effect of liraglutide on the browning of white adipocytes. CONCLUSION: Liraglutide induced browning of white adipose through regulation of miR-27b.


Assuntos
Liraglutida , MicroRNAs , Tecido Adiposo Marrom , Tecido Adiposo Branco , Animais , Dieta Hiperlipídica/efeitos adversos , Liraglutida/farmacologia , MicroRNAs/genética , Obesidade/tratamento farmacológico , Obesidade/genética , Ratos , Ratos Sprague-Dawley
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...