Upregulation of microRNA-93-5p/microRNA-4668-5p,and promoter methylation of matrix metalloproteinase-3 and interleukin-16 genes in Turkish patients with rheumatoid arthritis

Aim of the workTo investigate promoter methylation of matrix metalloproteinase-3 (MMP-3) and interleukin-16 (IL-16) genes with the expression of miRNA-93-5p and miRNA-4668-5p which target these genes in rheumatoid arthritis (RA), respectively.Patients and methodsThe study included 49 RA patients and 38 healthy controls. Promoter methylation of MMP-3 and IL-16 was analyzed by methylation-specific PCR. The expression of miRNA-93-5p and miRNA-4668-5p were determined. Disease activity score (DAS28) was assessed.ResultsThe 49 patients (38 female, 11 male) mean age was 50.4 ± 10.5 years and disease duration of 9.1 ± 7.4 years. The mean DAS28 was 3.9 ± 1.4. The MMP-3 gene methylation frequency was significantly lower in patients (n = 37;75.5%) compared to control (n = 37;97.4%) (p = 0.004) while they were comparable for IL-16 gene (n = 46;93.9% vs n = 37;97.4%)(p = 0.45). The relative normalized expression of miRNA-93-5p and miRNA-4668-5p were significantly increased (p < 0.001) in patients (2.28 ± 3.71 and 2.47 ± 4.17-fold) compared to controls (1.12 ± 0.18 and 1.28 ± 0.53-fold) and both tended to decrease with high disease activity (r =  ? 0.104, p = 0.52; r = ?0.24, p = 0.15, respectively). There was no significant difference of miRNA-93-5p (p = 0.45), and miRNA-4668-5p (p = 0.26) expressions between patients receiving treatment and those not. A negative correlation was observed between disease activity and change in expression levels of miRNA-93-5p (r = ?0.104, p = 0.52) and miRNA-4668-5p (r = ?0.24, p = 0.15). The ROC curve analysis of target miRNAs showed the diagnostic potential of miRNA-93-5p and miRNA-4668-5p (p = 0.003 and p < 0.001 respectively).ConclusionsThe methylation status of MMP-3 promoter and expression levels of miRNA-93-5p and miRNA-4668-5p could be useful biomarkers for the pathogenesis of RA and might reflect disease activity.     

MicroRNA在人结肠癌干细胞中的表达谱

目的:探讨微小RNA(microRNA,miRNA)在人结肠癌干细胞中的表达,为进一步研究miRNA调控结肠癌干细胞向结肠癌细胞分化的分子机制奠定基础.方法:应用miRNA表达谱芯片检测人结肠癌干细胞和已分化结肠癌细胞中miRNA的表达谱.利用实时定量PCR技术检测两种细胞中差异表达的miRNA,验证miRNA芯片结果的可靠性.应用软件对筛选出的显著差异表达miRNA的靶基因进行预测.结果:与已分化结肠癌细胞相比,人结肠癌干细胞中表达上调超过1.5倍的miRNA有35个.为:hsa-miR-192,hsa-miR-29b,hsa-miR-215,hsa-miR-194,hsa-miR-33a,hsa-miR-32等:表达下调超过1.5倍的miRNA有11个,为:hsa-miR-93,hsa-miR-1231,hsa-miR-524-3p,hsa-miR-886-3p等.PCR技术验证,与miRNA芯片结果相符合.表达显著上调miRNA的共同靶mRNA有:AFF2、MTF1、RUNDC2C和ZFHX4.表达显著下调miRNA的共同靶mRNA有:ONECUT2、SH3TC2、PTPRT、RNABP10、NR3C1、RGSL1、RNASEL和TANC2.结论:筛选出的差异表达miRNA可能参与结肠癌的发病.为该病诊治提供了新的思路.其共同靶基因可能具有重要的调控结肠癌干细胞生长和分化的作用.     

Screening and confirmation of microRNA markers for distinguishing between menstrual and peripheral blood

The identification of menstrual blood (MB) and peripheral blood (PB) left at a crime scene is crucial for crime reconstruction, especially in sexual assaults. MicroRNAs (miRNAs), a class of non-protein coding molecules, have been demonstrated to be a viable tool for body fluid identification in forensic casework. Several groups have searched for miRNAs that are specific to different body fluids. Blood has been studied the most extensively. However, menstrual blood was only involved in five studies, and the results confirming the presence of specific miRNAs could not be reproduced in other studies. In this study, we attempted to screen new markers that can differentiate between menstrual blood and peripheral blood by using Exiqon’s miRCURY™ LNA Array. Five miRNAs were selected based on the microarray results, namely, miR-141-3p, miR-373-3p, miR-497-5p, miR-143-5p, and miR-136-5p, whose expression levels exhibited 27.95-, 17.96-, 16.74-, 10.14-, and 9.21-fold changes, respectively, compared to the level in peripheral blood. Two classic quantitative methods, TaqMan hydrolysis probes (TaqMan for short) and SYBR Green fluorochrome (SYBR Green for short), were applied in the confirmation step to study the impact of different quantitative methods on the results. Three miRNAs (miR-141-3p, miR-497-5p, and miR-143-5p) were confirmed by TaqMan and one (miR-141-3p) by SYBR Green. Furthermore, bioinformatic methods were applied to interpret the candidate miRNAs. Our results established a multi-step procedure for body fluid identification and showed that the choice of quantitative method is important when miRNAs are used to identify the origin of blood samples.     

miRNA-21与心血管疾病

MicroRNAs (miRNAs)是长度约22个核苷酸左右的内源性、非编码RNA.在心血管疾病状态下,如增殖性血管疾病,心脏肥大,心功不全和缺血性心脏病的心血管组织中,miRNA-21表达下调.miRNA-21在血管平滑肌细胞的增殖、凋亡,心肌细胞生长、死亡及心肌细胞纤维化方面起着重要作用,已被证实参与上述疾病发病过...     

MicroRNA与肝癌的相关研究

肝癌是消化系统的常见恶性肿瘤,早期诊断困难,治疗手段有限,总体预后较差。近年来随着对MicroRNA深入研究,人们逐渐意识到MicroRNA与肝癌的发生、发展、转移等密切相关,并且在肝癌的早期诊断、治疗及预后评估中有着广阔的前景。本文就近年来关于MicroRNA与肝癌的相关研究作一概述。     

微小核糖核酸在参与缺血/再灌注过程中的研究进展

微小核糖核酸(miRNA)是一种近年来发现的存在于真核生物中的非编码调控RNA,其作用机制主要是在转录后水平上调控基因的表达。不同miRNA在炎性反应以及组织缺血/再灌注(I/R)过程中的表达水平有不同程度的变化,炎性反应是介导I/R损伤的重要因素之一,这一现象初步揭示了miRNA在I/R过程中扮演着重要的角色。深入研究将miRNA早期诊断和治疗I/R损伤有重要的临床意义。     

MicroRNA single-nucleotide polymorphisms and diabetes mellitus: A comprehensive review

Diabetes mellitus (DM) has become the third major chronic non-communicable disease affecting global public health, following cancer and cardiovascular and cerebrovascular diseases. Although previous studies have found a correlation between microRNA (miRNA) and the development of DM, thus far, most reviews have focused on the studies describing the changes in miRNA expression profiles and the mechanisms by which miRNAs-induce DM. However, reviews summarizing the effect of miRNA single-nucleotide polymorphisms on the developmental stages of DM and its complications are still needed. Studies on the variation of miRNAs will not only help to further understand the role that genetic factors play in DM, but will also have important implications for treatment and disease prognosis.     

MicroRNA-1、MicroRNA-133、MicroRNA-34a及其可能靶蛋白Ankyrin-B在心房颤动患者心房组织中的表达及意义

目的 通过检测心房颤动（房颤）与窦性心律患者右心耳组织中microRNA-1、microRNA-133及microRNA-34a的表达差异,及其可能靶蛋白锚蛋白-B （Ankyrin-B ）的表达变化,分析两者之间的关系,以从microRNAs调控水平研究房颤发生、发展的新机制。 方法 将第三军医大学新桥医院40例风湿性心瓣膜病行心瓣膜置换术患者分为房颤组［男8例,女12例;年龄（52.9±5.8）岁］和窦性心律组［男9例,女11例;年龄（52.4±6.2）岁］ 。采用逆转录-实时定量聚合酶链式反应（RTQ-PCR,Real-time quantitive PCR）法检测患者右心耳组织中microRNA-1、microRNA-133及microRNA-34a的表达,ΔΔCt法计算结果;石蜡切片免疫组织化学法检测组织中Ankyrin-B的表达;蛋白免疫印迹（Western Blotting）法检测组织中Ankyrin-B的表达变化。 结果 房颤组患者右心房组织中的microRNA-1表达较窦性心律组显著降低（0.559±0.252 vs. 3.997±1.251;t =-21.455,P=0.000）,microRNA-133的表达较窦性心律组显著降低（0.630±0.238 vs. 5.514±1.549;t =24.133,P=0.000）,而microRNA-34a的表达较窦性心律组增高（4.783±2.012 vs. 1.350±0.638,t =12.596,P=0.000）。免疫组织化学法及Western Blotting均显示房颤组心房组织Ankyrin-B表达较窦性心律组明显降低,且差异有统计学意义（0.66±0.45 vs. 1.09±0.42;t =-3.396,P=0.001）。结论 MicroRNA-34a可能通过调节Ankyrin-B蛋白的表达,从而参与心房颤动的发生、发展。