Role of microRNA-21 and microRNA-155 as biomarkers for bronchial asthma

MicroRNA (miRNA)-21 and miRNA-155 are important regulators of gene expression of different immunological molecules. This study aimed to investigate the role of miRNA-21 and miRNA-155 as biomarkers in asthma by comparing their serum expression levels in asthmatic patients to those in healthy controls and correlating their levels with serum IL-4. The expression levels of miRNA-21 and miRNA-155 were evaluated by quantitative RT-PCR. Serum levels of IL-4 were determined using ELISA. Asthmatic patients showed significantly higher serum miRNA-21 and miRNA-155 expression levels compared to controls. A statistically significant positive correlation between the expression levels of miRNA-21 and IL-4 serum levels in asthmatic patients was detected. Nonetheless, no correlation was detected between miRNA-155 expression and each of IL-4 and miRNA-21. A receiver operating characteristic curve analysis showed that at a cut-off value of 1.37, the sensitivity of miRNA-21 as an asthma biomarker was 100% and the specificity was 95%. At a cut-off value of 1.96, the sensitivity of miRNA-155 as an asthma biomarker was 100% and the specificity was 100%. It can be concluded that miRNA-21 and miRNA-155 are potential non-invasive biomarkers in the diagnosis of eosinophilic asthma and its response to therapy.     

Circulating microRNA-133, microRNA-17 and microRNA-25 in serum and its potential diagnostic value in gastric cancer

Gastric cancer is one of the most common malignancies in the world and is considered as the most lethal gastrointestinal cancer. microRNAs (miRNAs) can be very important in detecting a disease at an early stage. The aim of this study was to investigate the microRNA-17 (miR-17), miR-25, and miR-133b in the serum of gastric cancer subjects. Serum samples were obtained from 120 gastric cancers and 102 healthy subjects. We evaluated expression levels of miR-17, miR-25 and miR-133b by quantitative real-time polymerase chain reaction. Our results showed that in the patients with gastric cancer, the expression level of miR-17 and miR-25 were significantly increased compared with the control group (P < 0.5), while the expression level of miR-133b was significantly decreased in patient groups compared with control cases (P < 0.5). It seems that expression of miRNAs in Iranian patients with gastric cancer is similar to other patients in other populations. These findings suggested that miR-17, miR-25 and miR-133b could be introduced as potential diagnostic candidates for the detection in gastric cancer patients in the early stage.     

microRNA-342, microRNA-191 and microRNA-510 are differentially expressed in T regulatory cells of type 1 diabetic patients

Regulatory T cells (Tregs) are critical regulators of autoimmune diseases, including type 1 diabetes mellitus. It is hypothesised that Tregs’ function can be influenced by changes in the expression of specific microRNAs (miRNAs). Thus, we performed miRNAs profiling in a population of Tregs separated from peripheral blood of five type 1 diabetic patients and six healthy donors. For more detailed molecular characterisation of Tregs, we additionally compared miRNAs expression profiles of Tregs and conventional T cells. Tregs were isolated according to CD3+, CD4+, CD25^hi+ and CD127− by flow cytometry, and miRNA expression profiling was performed using TaqMan Array Human MicroRNA Panel-1 (384-well low density array). In Tregs of diabetic patients we found significantly increased expression of miRNA-510 (p = 0.05) and decreased expression of both miRNA-342 (p < 0.0001) and miRNA-191 (p = 0.0079). When comparing Tregs and T cells, we revealed that Tregs had significant higher expression of miRNA-146a and lower expression of eight specific miRNAs (20b, 31, 99a, 100, 125b, 151, 335, and 365). To our knowledge, this is the first study demonstrating changes in miRNA expression profiles occurring in Tregs of T1D patients and a miRNAs signature of adult Tregs.     

血清microRNA-152和microRNA-602检测在肝癌诊断及手术疗效评估中的应用

为探讨microRNA-152和microRNA-602检测在肝癌诊断及手术疗效评估中的应用价值。采用实时荧光定量PCR检测佳木斯市中心医院2012年3月~2015年3月的19例肝癌血清标本中microRNA-152和microRNA-602的表达水平,分析microRNA-152和microRNA-602在乙型肝炎病毒(HBV)阳性组、HBV阴性组和健康对照组血清样本中的表达差异。结果发现HBV阳性血清样本中microRNA-152的表达水平(0.65±0.29)明显低于健康组(1.21±0.32),microRNA-602在HBV阳性血清样本中的表达(0.63±0.31)明显高于健康组(0.44±0.15),且水平表达的差异具有统计学意义(p0.05)。可见血清样本中的microRNA-152和microRNA-602可以用于HBV阳性肝癌诊断的血清标记物,microRNA-152可以用于HBV阳性肝癌术后效果评价的血清标记物。     

Teratogen-induced alterations in microRNA-34, microRNA-125b and microRNA-155 expression: correlation with embryonic p53 genotype and limb phenotype

Background  

In a large number of studies, members of the microRNA (miRNA)-34 family such as miRNA-34a, miRNA-34b, miRNA-34c, as well as miRNA-125b and miRNA-155, have been shown to be regulators of apoptosis. The ability of these miRNAs to perform this function is mainly attributed to their ability to interact with the p53 tumor suppressor, which is a powerful regulator of the teratologic susceptibility of embryos. We chose to explore whether miRNA-34a/b/c, miRNA-125b and miRNA-155 may play a role in teratogenesis by using p53^+/- pregnant mice treated with cyclophosphamide (CP) as a model. We evaluated how CP-induced alterations in the expression of these miRNAs in the embryonic limbs correlate with embryonic p53 genotype and CP-induced limb phenotypes.     

Diagnostic and mechanistic values of microRNA-130a and microRNA-203 in patients with papillary thyroid carcinoma

This research was determined to unearth the diagnostic values and the effects of microRNA (miR)-130a and miR-203 on cell proliferation and apoptosis of papillary thyroid carcinoma (PTC). Expression of miR-130a and miR-203 were evaluated and were subjected to correlation analysis. The diagnostic values of miR-130a and miR-203 and their associations with clinicopathological characteristics of patients with PTC were measured. The expression levels of miR-130a and miR-203 in K1, IHH4, TPC-1, and BCPAP cells together with Nthy-ori 3-1 cells were measured. Cells were transfected with miR-130a mimics, miR-203 mimics, and coordinate of miR-130a mimics and miR-203 mimics. Cell growth, colony formation, and apoptosis were detected by cell counting kit-8 (CCK-8) assay, colony formation assay, and flow cytometry. PTC tissues had decreased miR-130a and miR-203 relative to adjacent normal tissues and normal thyroid tissue (both P < .05). miR-130a was in positive correlation with miR-203 (r = 0.754, P < .01). miR-130a was related with tumor infiltration and tumor stage while miR-203 was implicated in tumor stage and lymph-node metastasis. The area under the curve (AUC), sensitivity, as well as specificity for miR-130 in predicting PTC was 0.839, 74.5%, and 85.0% and those for miR-203 were 0.818, 73.7%, and 84.0%, respectively. PTC cells had lower expression of miR-130a and miR-203 than that in Nthy-ori 3-1 cells. After transfected miR-130a and miR-203 mimics in BCPAP and TPC-1 cells, both cells had increased miR-130a and miR-203, promoted cell apoptosis rate and decreased cell growth rate, and colony formation ability. After coordinately transfected with miR-130a mimics and miR-203 mimics, the cell growth and colony formation ability of PTC cells were restrained, and apoptosis of PTC cells was elevated (all P < .05). This study highlights that miR-130a and miR-203 have satisfactory diagnostic value in PTC and upregulated miR-130a and miR-203 can inhibit PTC cell growth and promote cell apoptosis.     

Human circulating microRNA-1 and microRNA-126 as potential novel indicators for acute myocardial infarction

Circulating miRNAs have been shown as promising biomarkers for various pathologic conditions. The aim of this study was to clarify that circulating miR-1 and miR-126 in human plasma might be useful as biomarkers in acute myocardial infarction (AMI). In our study, after pre-test, two candidate miRNAs were detected by using real-time RT-PCR. Cardiac troponin I (cTnI) concentrations were measured by ELISA assay in plasma from patients with AMI (n=17) and healthy subjects (n=25), simultaneously. Increased miR-1 and decreased miR-126 in plasma from patients with AMI after the onset of symptoms compared with healthy subjects were found. A remarkable finding in this study is that miR-1, miR-126 and cTnI expression levels exhibited the same trend. Our results suggest that the plasma concentrations of miR-1 and miR-126 may be useful indicators for AMI.     

Expression of microRNA-1, microRNA-133a and Hand2 protein in cultured embryonic rat cardiomyocytes

In this study, we investigated the expression of the pathway, SRF–microRNA-1/microRNA-133a–Hand2, in the Wistar rat embryonic ventricular cardiomyocytes under conventional monolayer culture. The morphological observation of the cultured cardiomyocytes and the mRNA expression levels of three vital constituent proteins, MLC-2v, N-cadherin, and connexin43, demonstrated the immaturity of these cultured cells, which was featured by less myofibril density, immature sarcomeric structure, and significantly lower mRNA expression of the three constituent proteins than those in neonatal ventricular samples. More importantly, results in this study suggest that the change of SRF–microRNA-1/microRNA-133a–Hand2 pathway results into the attenuation of the Hand2 repression in cultured cardiomyocytes. These outcomes are valuable to understand the cellular state as embryonic cardiomyocytes to be in vitro model and might be useful for the assessment of engineered cardiac tissue and cardiac differentiation of stem cells.     

IGF-1 deficiency resists cardiac hypertrophy and myocardial contractile dysfunction: role of microRNA-1 and microRNA-133a

This study was designed to examine the impact of insulin-like growth factor-1 (IGF-1) deficiency on abdominal aortic constriction (AAC)-induced cardiac geometric and functional changes with a focus on microRNA-1, 133a and 208, which are specially expressed in hearts and govern cardiac hypertrophy and stress-dependent cardiac growth. Liver-specific IGF-1-deficient (LID) and C57/BL6 mice were subject to AAC. Echocardiographic and cardiomyocyte function were assessed 4 wks later. Haematoxylin and eosin staining was used to monitor myocardial morphology. Western blot and real-time PCR were used to detect protein and miR expression, respectively. Neonatal rat cardiomyocytes (NRCMs) were transfected with miRs prior to IGF-1 exposure to initiate cell proliferation. Immunohistochemistry and [(3)H] Leucine incorporation were used to detect cell surface area and protein abundance. C57 mice subject to AAC displayed increased ventricular wall thickness, decreased left ventricular end diastolic and end systolic dimensions and elevated cardiomyocyte shortening capacity, all of which were attenuated in LID mice. In addition, IGF-1 deficiency mitigated AAC-induced increase in atrial natriuretic factor, GATA binding protein 4, glucose transporter 4 (GLUT4) and Akt phosphorylation. In contrast, neither AAC treatment nor IGF-1 deficiency affected glycogen synthase kinase 3b, mammalian target of rapamycin, the Glut-4 translocation mediator Akt substrate of 160 kD (AS160) and protein phosphatase. Levels of miR-1 and -133a (but not miR-208) were significantly attenuated by AAC in C57 but not LID mice. Transfection of miR-1 and -133a obliterated IGF-1-induced hypertrophic responses in NRCMs. Our data suggest that IGF-1 deficiency retards AAC-induced cardiac hypertrophic and contractile changes via alleviating down-regulation of miR-1 and miR-133a in response to left ventricular pressure overload.     

MicroRNA-9过表达载体的构建与鉴定

目的:MicroRNA(miRNA)是一类对基因表达起着转录后调控的小分子RNA,在正常发育和疾病发生过程中都发挥着重要的功能。其中,miR-9是一个序列高度保守的成员,在神经发育中调节神经干细胞的多种行为,包括分化、迁移等。但是,目前研究发现的miR-9的功能还存在一些相互矛盾和不清楚的地方,因此,我们拟构建miR-9的过表达载体,以便于对miR-9进一步的仔细研究。方法:我们采用分子克隆的常用方法,以pcDNA6.2-GW/miR为基础,构建出pcDNA6.2-GW/miR-9的表达质粒,分别用PCR、酶切和测序的方法验证质粒构建的正确性,并在Hela细胞和NIH3T3细胞中验证该质粒是否可以过表达miR-9小分子。结果:我们成功构建出正确的pc DNA6.2-GW/miR-9表达质粒,并且该质粒无论在人源的Hela细胞还是在小鼠源性的NIH3T3细胞中都可以过表达miR-9小分子。结论:构建了一个在不同种属间通用的miR-9过载体,为我们进一步研究miR-9的功能奠定了基础。     

动物microRNA-181的功能与应用前景

microRNA(miRNA)是一类短的、进化上高度保守的非蛋白编码RNA, 长度一般为17~25个核苷酸, 通过阻止靶mRNA的翻译或与之互补配对诱导靶基因降解来调控其表达。文章简要总结了microRNA-181(miR-181)在动物细胞增殖、凋亡和分化中的作用和调控机制, 探讨了miR-181对淋巴细胞的增生分化、自身免疫、炎症和抗病毒等方面的免疫调控作用, 并简要分析了miR-181在肿瘤发生发展、诊断、治疗和预后等方面的功能与价值, 最后对miR-181的应用前景进行了探讨。研究miR-181家族成员的功能对于理解生命活动机制、疾病发生发展和找到诊治相关疾病的新方法等都具有重要的意义。     

microRNA-378 与相关疾病的研究进展

microRNAs是一类内源性表达的、长度约为22个核苷酸的非蛋白编码的单链RNA分子,是重要的转录后基因表达调控因子。在多种生理和病理过程中发挥重要作用,到目前为止,在动植物以及病毒中已经发现有24521个miRNA分子,miR-378是其中的一种,miR-378通过多种机制与众多疾病的发生发展密切相关。miR-378在不同肿瘤组织中起到不同作用,在胃癌,肝癌,结直肠癌等肿瘤中起到抑癌基因的作用,在白血病,胰腺癌,卵巢癌等肿瘤中起到癌基因的作用。在心血管方面,miR-378可以通过多种机制起到保护血管,延缓心血管疾病的发展。在骨代谢方面,miR-378可通过不同机制抑制或促进成骨细胞的分化。本文就其与肿瘤、心血管、骨代谢以及其他方面的研究进行介绍,为这些疾病的治疗和预防提供一种新的思路。     

Sequence-specific inhibition of microRNA- and siRNA-induced RNA silencing

A large number of miRNAs have recently been discovered in plants and animals. Development of reverse genetic approaches that act to inhibit microRNA function would facilitate the study of this new class of noncoding RNA. Here we show that 2'-O-methyl oligoribonucleotides, but not 2'-deoxyoligonucleotides specifically inactivate the RNAi activity associated with miRNA-protein complexes in human cell extracts as well as in cultured human cells.     

Liver-specific microRNA-122: Biogenesis and function

microRNA-122 (miR-122) was one of the first examples of a tissue-specific miRNA. It is highly expressed in liver, where it constitutes 70% of the total miRNA pool. miR-122 expression is specific to the vertebrate lineage, where the sequence of the mature miRNA is completely conserved. miR-122 is a target for extensive study due to its association with cholesterol metabolism and hepatocellular carcinoma, and its important role in promoting hepatitis C virus (HCV) replication. This review will discuss the biogenesis and function of miR-122.     

Role of microRNA-155 in autoimmunity

MicroRNAs (miRNAs) have recently emerged as a major class of gene expression regulators linked to most biological functions. MiR-155 is encoded within a region known as B cell integration cluster (Bic) gene, identified originally as a frequent integration site for the avian leukosis virus. Disregulation of endogenous miR-155 has been implicated in the pathogenesis of human cancers. Recently, aberrant expression of miR-155 was observed in many autoimmune conditions, including rheumatoid arthritis (RA), multiple sclerosis (MS), and systemic lupus erythematosus (SLE). Moreover, functional analysis demonstrated that miR-155 has powerful regulatory potential in a wide variety of immune cells through targeting specific mRNAs. Since pathogenic immune cells play a pivotal role in pathogenesis of human autoimmune diseases, miR-155 might be a versatile therapeutic target. This review will discuss the current understandings for the role of miR-155 in autoimmunity.     

microRNA-155研究进展

microRNAs(miRNAs)是一类进化上保守的非编码单链小RNA,长19～24个核苷酸,能在转录后降解靶基因mRNA或抑制基因的翻译。microRNA-155(miR-155)具有多种生物学功能,它能影响造血细胞的分化,并在炎症反应、免疫反应中发挥重要作用。大量研究表明,miR-155在多种肿瘤细胞中过表达,推测其与肿瘤的发生发展密切相关。随着研究的深入,miR-155可能成为新的肿瘤标志物及肿瘤基因治疗的新靶点。