MicroRNA-539靶向膜相关的鸟苷酸激酶蛋白1抑制甲状腺癌细胞的侵袭与迁移

目的 观察microRNA-539（miR-539）的表达对甲状腺癌细胞侵袭与迁移的影响,探究miR-539与膜相关的鸟苷酸激酶蛋白1（CARMA1）在甲状腺癌中相互作用的机制。方法 构建miR-539与CARMA1高表达及低表达的细胞模型后通过transwell及MTT法检测两种不同的甲状腺癌细胞的侵袭、迁移及增殖能力。Western blot及RT-PCR检测细胞中蛋白及mRNA表达。结果  miR-539抑制甲状腺癌细胞的侵袭与迁移。通过荧光素酶报告检测发现miR-539通过靶向CARMA1的3’-UTR端从而起到抑制甲状腺癌中CARMA1的作用。进一步的研究证实,CARMA1可显著促进甲状腺癌细胞的侵袭与迁移。而调节CARMA1的表达,miR-539随之变化。研究还发现,与对照组比较,甲状腺癌中miR-539的表达量降低而CARMA1的表达量增加。结论  miR-539靶向CARMA1抑制甲状腺癌细胞的侵袭与迁移。

     

microRNA定量检测方法研究现状

MicroRNA(miRNA)是一类由18～24 nt核酸构成的非编码内源性小分子RNA,主要参与真核生物的细胞分化、增殖与凋亡等转录后水平的基因表达调控.研究发现,miRNA与肿瘤、代谢调控、疾病发生及诊断等方面有密切的联系,因此准确且灵敏地进行miRNA的定量检测是深入研究miRNA功能的前提.目前,miRNA定量检测原理主要基于核酸杂交或扩增,包括Northern印迹、微阵列芯片、实时定量PCR、滚环扩增等.本文通过对传统miRNA检测方法、高灵敏度新型miRNA检测方法进行阐述与分析,为筛选合适的miRNA定量检测方法提供参考.     

Can we increase the speed and efficacy of antidepressant treatments? Part II. Glutamatergic and RNA interference strategies

In the second part we focus on two treatment strategies that may overcome the main limitations of current antidepressant drugs. First, we review the experimental and clinical evidence supporting the use of glutamatergic drugs as fast-acting antidepressants. Secondly, we review the involvement of microRNAs (miRNAs) in the pathophysiology of major depressive disorder (MDD) and the use of small RNAs (e.g.., small interfering RNAs or siRNAs) to knockdown genes in monoaminergic and non-monoaminergic neurons and induce antidepressant-like responses in experimental animals.The development of glutamatergic agents is a promising venue for antidepressant drug development, given the antidepressant properties of the non-competitive NMDA receptor antagonist ketamine. Its unique properties appear to result from the activation of AMPA receptors by a metabolite [(2 S,6 S;2 R,6 R)-hydroxynorketamine (HNK)] and mTOR signaling. These effects increase synaptogenesis in prefrontal cortical pyramidal neurons and enhance serotonergic neurotransmission via descending inputs to the raphe nuclei. This view is supported by the cancellation of ketamine's antidepressant-like effects by inhibition of serotonin synthesis.We also review existing evidence supporting the involvement of miRNAs in MDD and the preclinical use of RNA interference (RNAi) strategies to target genes involved in antidepressant response. Many miRNAs have been associated to MDD, some of which e.g., miR-135 targets genes involved in antidepressant actions. Likewise, SSRI-conjugated siRNA evokes faster and/or more effective antidepressant-like responses. Intranasal application of sertraline-conjugated siRNAs directed to 5-HT_1A receptors and SERT evoked much faster changes of pre- and postsynaptic antidepressant markers than those produced by fluoxetine.     

微RNA-196a和微RNA-210在胰腺癌中的表达及其与生存时间的关系

目的分析微RNA-196a(miR-196a)和miR-210在胰腺癌中的表达,及其与生存时间的关系。方法选择26例胰腺癌患者(A组)、20例良性胰腺病患者(B组)和24例健康受试者(C组)为研究对象。用实时定量荧光聚合酶链式反应法检测3组患者血清中miR-196a和miR-210表达水平。分析3组血清miR-196a和miR-210表达差异,miR-196a和miR-210水平与胰腺癌患者生存时间的关系。结果A,B,C组的miR-196a表达水平分别为(1.79±0.95),(1.24±0.92)和(0.71±0.68),miR-210表达水平(2.32±0.54),(1.25±0.36)和(0.98±0.22),A组的上述指标与B,C组比较,差异均统计学意义(均P<0.05)。在胰腺癌患者中,半年生存率为61.54%,1年生存率为46.15%,2年生存率为3.85%。在胰腺癌患者中,miR-196a高表达组和低表达组的生存时间分别为(7.58±3.43)和(12.38±5.17)个月,miR-210高表达组和低表达组的生存时间分别为(6.92±3.01)和(10.88±4.28)个月,差异均有统计学意义(均P<0.05)。miR-196a的敏感度、特异性及准确率分别为76.92%,93.18%和70.10%,miR-210的敏感度、特异性及准确率分别为68.00%,93.18%和61.18%,联合检测的敏感度、特异性及准确率分别为88.46%,95.45%和83.92%,联合检测的上述指标与miR-196a、miR-210单独检测比较,差异均有统计学意义(均P<0.01)。结论胰腺癌患者血清中特异性的表达miR-196a和miR-210,二者联合检测诊断胰腺癌可提高其诊断准确率,且miR-196a、miR-210表达水平与胰腺癌患者生存时间有一定关系。     

长链非编码RNA与微小RNA相互作用对调控疾病的研究进展

非编码RNA(ncRNA)与基因调控密切相关,ncRNA主要包括微小RNA(miRNA)和长链非编码RNA(lncRNA)。lncRNA与miRNA之间存在着相互调控关系,两者间的相互作用参与了众多疾病的发生发展。本文主要综述其在癌症、心血管疾病、免疫系统和神经系统疾病中的作用及其作用机制。     

Depressive disorder and antidepressants from an epigenetic point of view

Depressive disorder is a complex, heterogeneous disease that affects approximately 280 million people worldwide. Environmental, genetic, and neurobiological factors contribute to the depressive state. Since the nervous system is susceptible to shifts in activity of epigenetic modifiers, these allow for significant plasticity and response to rapid changes in the environment. Among the most studied epigenetic modifications in depressive disorder is DNA methylation, with findings centered on the brain-derived neurotrophic factor gene, the glucocorticoid receptor gene, and the serotonin transporter gene. In order to identify biomarkers that would be useful in clinical settings, for diagnosis and for treatment response, further research on antidepressants and alterations they cause in the epigenetic landscape throughout the genome is needed. Studies on cornerstone antidepressants, such as selective serotonin reuptake inhibitors, selective serotonin and norepinephrine reuptake inhibitors, norepinephrine, and dopamine reuptake inhibitors and their effects on depressive disorder are available, but systematic conclusions on their effects are still hard to draw due to the highly heterogeneous nature of the studies. In addition, two novel drugs, ketamine and esketamine, are being investigated particularly in association with treatment of resistant depression, which is one of the hot topics of contemporary research and the field of precision psychiatry.     

MicroRNA Profiling Reveals Distinct Signatures in Degenerative Rotator Cuff Pathologies

MicroRNAs (miRNAs) have emerged as key regulators orchestrating a wide range of inflammatory and fibrotic diseases. However, the role of miRNAs in degenerative shoulder joint disorders is poorly understood. The aim of this explorative case-control study was to identify pathology-related, circulating miRNAs in patients with chronic rotator cuff tendinopathy and degenerative rotator cuff tears (RCT). In 2017, 15 patients were prospectively enrolled and assigned to three groups based on the diagnosed pathology: (i) no shoulder pathology, (ii) chronic rotator cuff tendinopathy, and (iii) degenerative RCTs. In total, 14 patients were included. Venous blood samples (“liquid biopsies”) were collected from each patient and serum levels of 187 miRNAs were determined. Subsequently, the change in expression of nine candidate miRNAs was verified in tendon biopsy samples, collected from patients who underwent arthroscopic shoulder surgery between 2015 and 2018. Overall, we identified several miRNAs to be progressively deregulated in sera from patients with either chronic rotator cuff tendinopathy or degenerative RCTs. Importantly, for the several of these miRNAs candidates repression was also evident in tendon biopsies harvested from patients who were treated for a supraspinatus tendon tear. As similar expression profiles were determined for tendon samples, the newly identified systemic miRNA signature has potential as novel diagnostic or prognostic biomarkers for degenerative rotator cuff pathologies. © 2019 The Authors. Journal of Orthopaedic Research^® published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. Inc. J Orthop Res 38:202–211, 2020