类风湿关节炎患者外周血miR-146a、miR-155、Ets-1以及IRAK1的表达及临床意义

为探讨类风湿关节炎(rheumatoid arthritis,RA)患者PBMC中miR-146a、miR-155、Ets-1和IRAK1的表达水平及与病情活动度的相关性,收集RA高活动期患者23例、低活动期患者21例和健康志愿者22例,采用qRT-PCR检测PBMC中miR-146a、miR-155、Ets-1、IRAK1的相对表达水平,同时分析其与临床及实验室指标的相关性。RA患者PBMC的miR-146a、miR-155相对表达量显著高于健康对照组(P<0.05),高活动组高于低活动组(P<0.05)、低活动组与健康对照组差异无统计学意义(P>0.05);RA患者PBMC的Ets-1表达水平与健康对照组差异无统计学意义,高活动组显著低于低活动组和健康对照组(P<0.05)、低活动组与健康对照组差异无统计学意义(P>0.05);RA患者PBMC的IRAK1表达水平显著低于健康对照组(P<0.05),高活动组显著低于健康对照组(P<0.01)、低活动组显著低于健康对照组(P<0.05),而高活动组与低活动组间无统计学差异(P>0.05)。在RA患者中miR-155表达水平与血沉正相关(r=0.319,P=0.042)而与血红蛋白负相关(r=-0.386,P=0.017);Ets-1与CRP负相关(r=-0.408,P=0.007);IRAK1与RF、DAS 28、总补体负相关(r=-0.513,P=0.001;r=-0.332,P=0.029;r=-0.49,P=0.015)。研究结果显示,RA高活动组患者PBMC中miR-146a、miR-155表达升高,Ets-1、IRAK1表达降低,可能参与了RA发病过程的调控;但本研究样本量有限,miR-146a、miR-155和Ets-1、IRAK1参与RA发病的关系有待后续大样本研究证实,且相关调控机制仍需进一步深入研究。     

Pre-miR-146a基因rs2910164位点单核苷酸基因多态性及miR-146a表达与类风湿关节炎相关性研究

目的 探讨pre-miR-146a基因rs2910164位点单核苷酸基因多态性及miR-146a表达与类风湿关节炎相关性.方法 采用聚合酶链反应-连接酶检测反应检测123例类风湿关节炎(RA)患者和220例健康对照者pre-miR-146a rs2910164位点基因多态性,应用实时荧光定量聚合酶链反应检测68例RA患者、10例骨关节炎(OA)患者及20例健康对照外周血单个核细胞中miR-146a的表达水平,并选取10例RA疾病活动患者行激素加免疫抑制剂正规治疗3个月后miR-146a表达水平的测定.收集并计算RA患者临床参数:发病年龄、性别、类风湿因子(RF)和抗环瓜氨酸肽(抗-CCP)抗体、RA疾病活动(DAS28≥3.2)、骨破坏(X＞Ⅰ期).统计学处理采用X2检验、方差分析、t检验和Pearson相关分析.结果 RA组pre-miR-146a rs2910164位点的基因型频率和等位基因频率与健康对照组比较,差异无统计学意义(P均＞0.05).RA患者pre-miR-146ars2910164位点基因型与发病年龄、性别、RF和抗-CCP抗体阳性率、RA疾病活动、骨破坏阳性率及miR-146a表达量均无相关性(P均＞0.05).RA患者组miR-146a的表达量高于健康对照组和OA组(P均＜0.01),后两组miR-146a的表达量无统计学差异(P＞0.05).RA疾病活动组miR-146a表达高于非活动组和对照组(P均＜0.01),后两组miR-146a的表达量无统计学差异(P＞0.05).RA疾病活动患者治疗后miR-146a表达下降(P＜0.05),DAS28评分降低(P＜0.01).RA患者组miR-146a的表达与红细胞沉降率(ESR,即血沉)、C反应蛋白(CRP)及DAS28评分之间呈正相关(P均＜0.01),与RF、抗-CCP抗体滴度无相关性(P均＞0.05).结论 我国汉族人群中,pre-miR-146a rs2910164位点多态性与RA的易感性、临床参数及miR-146a的表达无相关性,RA患者外周血单个核细胞miR-146a表达上调,其表达水平与RA病情活动有关,miR-146a的检测可能是RA病情活动的一个有用的判断指标.     

miR-146a调控血小板免疫活化功能

目的探讨miR-146a对血小板免疫活化功能的影响。方法流式细胞术检测冠心病(n=31)及健康成年人(n=35)全血中血小板PAC-1、CD62-P阳性率,q-PCR及光比浊法分别检测血浆中miR-146a及血小板聚集率;培养K562细胞,佛波酯(PMA)诱导分化为巨核细胞后,分别转染miR-146a mimics、inhibitor及其阴性对照,Western blot检测转染后细胞中TLR-4、PAC-1及CD62-P表达水平。结果冠心病患者miR-146a、血小板聚集率及PAC-1、CD62-P阳性率较对照组升高(P0.05);转染miR-146a mimics后,TLR-4、PAC-1和CD62-P表达水平升高(P0.05)。结论miR-146a可能依赖血小板TLR-4信号通路,介导血小板免疫活化进而促进血小板聚集,加速血栓形成。     

miR-23a/b在非小细胞肺癌的表达及临床意义

目的研究miR-23a和miR-23b在非小细胞肺癌中的表达特征,并探讨其临床意义。方法收集157例非小细胞肺癌手术切除标本及50例癌旁组织标本,采用Real time PCR方法检测miR-23a和miR-23b在非小细胞肺癌及其癌旁组织中的表达,分析二者表达的相关性,并探讨其表达与临床病理特征及其预后的关系。结果 miR-23a和miR-23b在非小细胞肺癌组织中的表达水平均高于癌旁肺组织,二者在非小细胞肺癌组织中表达呈正相关(r=0.351,P0.001)。miR-23a和miR-23b联合高表达与淋巴结有无转移(P0.001)、远处转移(P=0.001)及临床分期(P=0.002)相关,而与年龄、性别、组织类型及组织分化程度无显著相关性(P0.05)。KaplanMeier分析显示miR-23a和miR-23b联合高表达组患者生存期显著低于单独高表达组或联合低表达组(P=0.009),Cox风险比例模型分析显示miR-23a和miR-23b联合高表达为非小细胞肺癌患者的危险因素。结论 miR-23a和miR-23b在非小细胞肺癌中异常高表达可能是潜在的肺癌预后分子标志物。     

系统性红斑狼疮患者中miR-146a表达缺陷与I型干扰素通路过度活化相关

Ι型干扰素(IFN)通路过度活化与固有免疫反应异常在系统性红斑狼疮(SLE)发病中起重要作用。miR-146a作为一个负调控因子,在固有免疫应答中发挥重要作用。为探索miR-146a在SLE发病中的作用,通过实时荧光定量聚合酶链反应技术检测18例SLE患者和11例正常对照miR-146a的表达,发现同正常对照组相比,miR-146a表达水平在SLE患者中明显降低,SLE活动组中下降则更加明显;同时检测miR-146a的靶基因IRAK1和TRAF6 mRNA表达,发现在SLE患者中其水平显著高于正常对照组;进一步分析miR-146a与IFN诱导基因表达之间的关系,结果显示两者之间存在负相关,体外实验也证实miR-146a能够负向调节I型干扰素通路。综上所述,我们的研究结果提示miR-146a可能作为一个新的生物标志物,SLE患者中miR-146a表达水平下降,导致I型干扰素通路负调控作用被削弱,从而参与疾病的发生发展。     

类风湿关节炎患者外周血miR-146a、miR-16的表达水平及临床指标与中医证型的关系探讨

目的研究类风湿关节炎（RA）患者外周血miR-146a、miR-16的表达及ESR、CRP、RF、抗CCP抗体与中医证型的关系,为RA辨证分型提供客观依据。方法根据《中药新药临床研究指导原则》有关RA的中医证候诊断标准,主要分为两个证型：风湿夹瘀型（n=20）和肝肾亏损型（n=16）;16名健康体检者作为健康对照组。Real-timePCR检测RA患者及健康对照组miR-146a、miR-16的表达,记录ESR、CRP、RF、抗CCP抗体等临床指标。结果风湿夹瘀型miR-146a、miR-16的表达水平高于肝肾亏损型和对照组（P均〈0.05）,肝肾亏损型miR-146a、miR-16的表达水平与健康对照组相比差异无统计学意义（P均〉0.05）,风湿夹瘀型RA患者ESR及CRP高于肝肾亏损型（P〈0.01和P〈0.05）,RF低于肝肾亏损型（P〈0.05）,抗CCP抗体在两种证型之间差异无统计学意义（P〉0.05）。结论 miR-146a、miR-16的表达水平和ESR、CRP、RF等临床指标可能为RA风湿夹瘀及肝肾亏损型的辨证分型提供参考。     

血清miR-146a变化与强直性脊柱炎的相关性

目的:探究强直性脊柱炎血清miR-146a表达情况,从而发现潜在的强直性脊柱炎的生物标记分子。方法:qRTPCR检测强直性脊柱炎患者和正常人血清miR-146a表达水平。根据脊柱后凸的程度将患者分为A(后凸畸形70%)和B(后凸畸形≥70%)两组。疾病的活性以及患者功能状态分别由强直性脊椎炎疾病活动度指数(BASDAI)和强直性脊椎炎功能指数(BASFI)来表示。采用qRT-PCR检测A和B两组血清miR-146a表达水平,绘制受试者工作特征(ROC)曲线评价血清中miR-146a对强直性脊柱炎的诊断价值,同时将miR-146a表达量与临床指标进行相关性分析。结果:miR-146a在强直性脊柱炎血清组表达明显上调(P0.05)。miR-146a的ROC曲线下面积值为0.917;且B组强直性脊柱炎患者血清中miR-146a表达水平显著高于A组(P0.05)。此外,miR-146a表达水平同BASDAI之间存在显著相关性(r=0.5,P0.05)。结论:血清中miR-146a表达检测可作为强直性脊柱炎诊断的辅助性生物标记。miR-146a表达水平也可能与疾病活性和胸腰椎后凸畸形的严重程度成一定的相关性。     

miR-132和miR-146a在内侧颞叶癫痫患儿和幼年大鼠海马组织中的表达变化

目的:观察miR-132、miR-146a在内侧颞叶癫痫(mesial temporal lobe epilepsy,MTLE)患儿及幼年大鼠模型海马组织中的表达变化。方法:收集MTLE患儿海马标本及正常对照人海马标本,Real-time PCR方法检测两组人海马组织中miR-132、miR-146a的表达改变。利用匹罗卡品诱导建立幼年大鼠MTLE模型,收集急性期(造模后2 h)、潜伏期(造模后3周)、慢性期(造模后8周)海马组织标本及相应时间点对照组大鼠海马标本,Real-time PCR方法检测各实验组大鼠海马组织中miR-132、miR-146a的表达改变。结果:miR-132、miR-146a在患儿MTLE海马组织中表达明显升高,分别为2.2±0.1和3.0±0.2,与正常对照组(1.0±0.2)比较差异有统计学意义(P0.05)。miR-132在MTLE大鼠模型急性期、潜伏期及慢性期表达均明显增加,分别为5.2±0.5,2.5±0.2和3.6±0.2,较正常对照组大鼠(1.0±0.1)差异有显著性(P0.05);miR-146a在MTLE大鼠急性期改变不明显,在潜伏期及慢性期表达明显增加(2.8±0.2,1.8±0.2),与对照组大鼠(1.0±0.1)比较差异有统计学意义(P0.05)。结论:miR-132、miR-146a在MTLE患儿及幼年大鼠模型中存在差异表达,其可能与MTLE中脑组织炎症反应的异常调控有关。     

中国广西人群miR-146a、miR-149 基因遗传多态性的研究

目的：研究广西人群miR-146a C>G (rs2910164）、miR-149 T>C（rs2292832）等位基因及基因型频率分布,分析其与不同民族种族间的差异性。方法：采用单碱基延伸技术和DNA测序技术对303例广西人群中miR-146a C>G和miR-149 T>C基因单核苷酸多态性(SNP)位点进行分型检测,并与人类基因组计划 (Hapmap) 数据库中公布的非洲人、欧洲人、日本人和中国北京人群的SNP分型数据比较。结果：miR-146a C>G、miR-149 T>C等位基因和基因型频率在广西男女人群之间分布均无差异性（P均>0.05）。广西人群miR-146a C>G和miR-149 T>C基因型及等位基因频率与非洲人、欧洲人、中国北京人比较有统计学意义（P均<0.05）。结论：广西人群miR-146a C>G和miR-149 T>C存在基因多态性,与其他种族人群比较有差异性,这种差异性对人类遗传病的研究可能会起到重要作用。     

乳腺癌中miR-146a对肿瘤相关巨噬细胞功能的影响

目的探讨miR-146a对肿瘤相关巨噬细胞(TAM)功能的影响及其在TAM中表达水平变化的调控机制。方法分别对比BALB/c小鼠4T1移植瘤TAM和腹腔巨噬细胞(PEC)、人乳腺癌组织TAM和外周血单核细胞,q-PCR法检测miR-146a的表达变化。转染miR-146a antagomir的巨噬细胞与4T1混合接种,确定TAM中miR-146a表达对肿瘤发生发展的影响。结果 miR-146a在荷瘤小鼠(P0.05)和人乳腺癌(P0.01)相关巨噬细胞中显著下调。miR-146a inhibitor明显抑制肿瘤的生长。结论 TAM中miR-146a可能通过其对巨噬细胞的调控进而促进肿瘤生长。     

MiR-146与免疫及自身免疫性疾病

MicroRNAs (miRNAs)介导的基因表达对于维持正常的细胞周期、分化、增殖、凋亡以及维持免疫系统的稳定性方面发挥着非常重要作用.MiR-146家族包含miR-146a和miR-146b,在不同的造血细胞中具有不同的表达谱.研究发现miR-146与炎症、自身免疫性疾病密切相关,负性调节炎症和固有免疫反应,可作为...     

MiR-146a modulates macrophage polarization in systemic juvenile idiopathic arthritis by targeting INHBA

Monocytes from patients with systemic juvenile idiopathic arthritis (SJIA) have both features of classical activated M1 and alternatively activated M2 macrophages. An increasing number of studies have indicated that microRNAs (miRNAs) are critical regulators of monocyte polarization. Here, we focused on miR-146a expression in SJIA and investigated the function of miR-146a in monocyte polarization. We found that miR-146a expression was highly up-regulated in SJIA monocytes and correlated with the systemic features. miR-146a was expressed at a higher level in monocytes polarized with M2 conditions than those polarized with M1 conditions. miR-146a overexpression significantly decreased the production of M1 phenotype markers such as IL-6, IL-12, TNF-α, CD86 and iNOS in M1 macrophages, but increased the production of M2 marker genes such as Arg1, CCL17, CCL22 and CD206 in M2 macrophages. Conversely, knockdown of miR-146a promoted M1 macrophage polarization but diminished M2 macrophage polarization. We subsequently demonstrated that miR-146a targeted the 3′-untranslated region (UTR) of INHBA to inhibit its expression. Additionally, INHBA overexpression rescued the reduced IL-6, IL-12, and TNF-α levels induced by miR-146a overexpression in M1 macrophages, and rescued the increased Arg1, CCL17, and CCL22 levels induced by miR-146a overexpression in M2 macrophages. Similarly, the effects of miR-146a inhibition in monocyte polarization were all partly reversed by INHBA inhibition. Taken together, the data suggest that miR-146a serves as a molecular regulator in monocyte polarization and might play an important role in monocytes from patients with SJIA.     

MiR-146a and miR-155 polymorphisms in Egyptian patients with Behcet’s disease

IntroductionThe current study was designed to analyze whether polymorphisms of miR-146a and miR-155 are related to Behçet’s disease (BD) in the Egyptian population.Material and methodsA total of 96 unrelated BD patients and 100 healthy subjects were genotyped for miR-146a (rs2910164) and miR-155 (rs767649) using real-time polymerase chain reaction.ResultsThe results showed significant elevation in the frequency of rs2910164 GG and CC genotypes in BD patients compared with controls (adjusted OR = 22.156, 95% CI: 4.728–103.818; p < 0.001 and adjusted OR = 40.358, 95% CI: 8.928–182.440; p < 0.001, respectively). Also, the rs2910164 G allele conferred a higher risk of developing BD (adjusted OR = 3.665, 95% CI: 2.013–6.671; p < 0.001). MiR-146a (rs2910164) polymorphism was a risk factor for susceptibility to BD in dominant, recessive and additive models of inheritance (all p < 0.001), while the miR-155 (rs767649) polymorphism was a risk factor in the recessive model only (p = 0.021). GG and CG genotypes of rs2910164 were associated with higher Behcet’s disease current activity index (BDCAI) and ocular involvement compared with CC genotype (p = 0.005 and p = 0.004, respectively). Genotype AT of rs767649 was related to higher BDCAI (p = 0.026) compared with TT and AA genotypes.ConclusionsmiR-146a (rs2910164) and miR-155 (rs767649) are likely to play an important role in the Egyptian population in development of BD and also influence disease severity.     

miR-144、miR-451在胃癌组织中的表达及其临床意义

目的探究胃癌患者组织中miR-144、miR-451的表达与临床病理参数及预后的关系。方法2013年6月至2014年6月,收集115例胃癌患者的癌组织及癌旁正常组织,采用实时定量PCR检测组织中miR-144、miR-451的表达水平;分析胃癌组织中miR-144、miR-451的表达与患者临床病理参数的关系,采用Kaplan-Meier生存曲线评估miR-144、miR-451的表达与患者预后的关系,并采用Cox回归模型分析胃癌患者预后的影响因素。结果胃癌组织中miR-144相对表达量为0.214±0.069,低于癌旁正常组织的1.124±0.157,miR-451相对表达量为0.354±0.087,低于癌旁正常组织的1.084±0.108,差异均有统计学意义(均P<0.05);miR-144、miR-451的表达均与肿瘤大小、淋巴结转移、远处转移及TNM分期有关(均P<0.05)。Kaplan-Meier生存分析结果表明,与miR-144高表达组、miR-451高表达组相比,miR-144低表达组、miR-451低表达组患者术后5年总生存率较低,术后中位生存时间较短(均P<0.05)。Cox回归模型发现,远处转移、较高TNM分期、miR-144低表达、miR-451低表达是影响胃癌患者预后的危险因素(均P<0.05)。结论胃癌组织中miR-144、miR-451呈低表达,其与患者病情进展和预后有关,检测胃癌组织中miR-144、miR-451的表达可能有利于患者的病情和预后判断。     

The microRNAs, MiR-31 and MiR-375, as candidate markers in Barrett's esophageal carcinogenesis

There is a critical need to identify molecular markers that can reliably aid in stratifying esophageal adenocarcinoma (EAC) risk in patients with Barrett's esophagus. MicroRNAs (miRNA/miR) are one such class of biomolecules. In the present cross-sectional study, we characterized miRNA alterations in progressive stages of neoplastic development, i.e., metaplasia-dysplasia-adenocarcinoma, with an aim to identify candidate miRNAs potentially associated with progression. Using next generation sequencing (NGS) as an agnostic discovery platform, followed by quantitative real-time PCR (qPCR) validation in a total of 20 EACs, we identified 26 miRNAs that are highly and frequently deregulated in EACs (≥ 4-fold in >50% of cases) when compared to paired normal esophageal squamous (nSQ) tissue. We then assessed the 26 EAC-derived miRNAs in laser microdissected biopsy pairs of Barrett's metaplasia (BM)/nSQ (n = 15), and high-grade dysplasia (HGD)/nSQ (n = 14) by qPCR, to map the timing of deregulation during progression from BM to HGD and to EAC. We found that 23 of the 26 candidate miRNAs were deregulated at the earliest step, BM, and therefore noninformative as molecular markers of progression. Two miRNAs, miR-31 and -31*, however, showed frequent downregulation only in HGD and EAC cases suggesting association with transition from BM to HGD. A third miRNA, miR-375, showed marked downregulation exclusively in EACs and in none of the BM or HGD lesions, suggesting its association with progression to invasive carcinoma. Taken together, we propose miR-31 and -375 as novel candidate microRNAs specifically associated with early- and late-stage malignant progression, respectively, in Barrett's esophagus.