多发性骨髓瘤50例患者miRNA-21、 miRNA-29、 miRNA-15a和 miRNA-16水平

摘 要：［目的］ 探讨miRNAs在多发性骨髓瘤（multiple myeloma,MM）的临床价值。［方法］ 选择2014年1月至2016年1月于宝鸡市中医医院收治的多发性骨髓瘤患者50例,分别取其化疗前后的血清作为病例组,另选择同期体检的50名健康志愿者血清作为对照组。分别检测受试者血清以及病例组患者化疗前后血清中miRNAs的表达含量。［结果］ 病例组患者的miRNA-21和miRNA-29的表达水平显著性高于对照组（P<0.05）,miRNA-15a以及miRNA-16等含量显著性低于对照组（P<0.05）。MM患者经化疗后,血清miRNA-21以及miRNA-29显著性降低（P<0.05）,miRNA-15a和miRNA16的含量显著性升高（P<0.05）,且化疗疗效有效的患者血清中相应miRNA指标变化水平较化疗疗效无效患者明显（P<0.05）。［结论］　miRNAs可能参与多发性骨髓瘤的发生发展,其异常表达情况可能为该疾病的预测和预后评价提供临床价值。     

microRNAs在胃癌发生发展中的研究进展

微小RNAs（miRNAs）是一组小的非编码RNA,在胃癌中起着至关重要的作用。miRNAs通过靶基因mRNA产生双重的、相反的作用,即致癌或抑癌作用。致癌miRNAs在胃癌中过表达,并能抑制肿瘤抑制基因。相反,肿瘤抑制miRNAs在胃癌中表达通常下调,从而导致癌症的发展。异常表达的miRNAs参与胃癌的发生发展并调控不同的表型,如增殖、凋亡、转移及耐药性等。此外,miRNAs还可用于胃癌的诊断和预后预测等。本文综述了miRNAs在胃癌发展中的作用,以及miRNAs在诊断和预后中的潜在应用价值。     

小鼠支气管肺泡干细胞miRNAs表达谱的鉴定

[目的]检测并验证正常成年小鼠肺内支气管肺泡干细胞（BASCs）的miRNAs表达情况。[方法]流式细胞仪分选小鼠BASCs和其对照细胞（CD45-CD31-Sca-1-CD34-细胞）;将分选出的细胞常规提取RNA后,经YM-100（Millipore）微离心过滤柱抽提小于300核苷酸长度的小RNA;微阵列法检测BASCs和其对照细胞的miRNAs表达谱,筛选出差异miRNAs;构建miRNAs特异性TaqManMGB探针,qRT-PCR法验证所选差异miRNAs在两种细胞的表达。[结果]微阵列法检测显示BASCs和对照细胞有116个miRNAs的表达有显著性差异（P〈0.01）,其中有56个miRNAs在BASCs高表达,60个miRNAs在BASCs低表达。在这些差异miRNAs中选取了10个与细胞周期、干细胞分化、肿瘤发生相关的miRNAs,通过qRT-PCR法检测其在BASCs和对照细胞中的表达,qRT-PCR的检测结果与miRNAs芯片检测结果一致。[结论]研究通过微阵列法鉴定了小鼠BASCs的miRNAs表达谱,与对照细胞相比,发现116个miRNAs的表达具有显著性差异（56个高表达,60个低表达）。     

微小RNA研究进展

微小RNA(miRNA)是参与基因转录后水平调控的非编码小分子RNA.人类基因中大约有3%编码miRNAs,而编码蛋白的基因中30%受到miRNAs的调控.miRNAs在多种生物进程中起到关键作用,包括调节发育、细胞增殖、分化和凋亡,相应的miRNAs的表达变化与包括肿瘤在内的多种疾病有关.本文综述miRNAs的生物学及其与肿瘤的联系,并讨论了miRNAs的研究方法.     

外泌体microRNAs在卵巢癌诊断和治疗中作用的研究进展

外泌体microRNAs(miRNAs)在卵巢癌进展的各种过程中发挥重要作用，如卵巢癌细胞增殖、凋亡和侵袭。外泌体miRNAs通过各种途径参与基因表达、信使RNA合成、蛋白生成等，有助于对卵巢癌发病机制的理解。由于miRNAs在血浆外泌体中的有效性，miRNAs可作为新兴的癌症早期检测和治疗评估生物标志物。外泌体miRNAs作为一种信使，在肿瘤微环境中起着重要的作用，越来越多的研究也证实了miRNAs作为靶点治疗卵巢癌的可行性。     

微小RNAs在良恶性胸腔积液中的研究进展

microRNAs(miRNAs）是长度为19~24 nt的非编码小RNA,可通过与其靶mRNA特异性结合导致mRNA降解或翻译抑制,从而负向调控基因表达。miRNAs参与包括细胞分化、增殖、凋亡等一系列重要细胞活动的调控。miRNAs的异常表达与人类肿瘤密切相关。miRNAs具有较强的稳定性,广泛存在人体组织、体液中。本文就miRNAs在胸腔积液中的研究进展作一综述,以期为良恶性胸腔积液的鉴别等提供新的概念及思路。     

结直肠癌中microRNA标志物临床诊断价值的研究进展

结直肠癌是最常见的消化道恶性肿瘤,其发生、发展及诊疗、预后已成为当前临床研究关注 的热点。microRNAs (miRNAs) 是一类内源性的、19~25个碱基长度的小分子非编码RNA,在人体生命 活动中具有广泛的调节功能。目前发现,部分miRNAs异常表达与结直肠癌的发生、发展密切相关,提 示miRNAs可作为结直肠癌临床早期诊断及预后评估的特异性新型生物标志物。文章综述了miRNAs作 为结直肠癌临床早期诊断、预后和治疗结果评估生物标志物的研究进展,揭示结直肠癌miRNAs参与的 基因表达调控机制, 展望miRNAs作为结直肠癌新型诊断分子标志物及治疗靶点的广阔临床应用前景。     

长期高剂量苯并(a)芘染毒对小鼠肺脏microRNAs表达的影响

目的:研究长期高剂量苯并(a)芘[B(a)P染毒对小鼠肺组织miRNAs表达谱的影响,探讨miRNAs在B(a)P健康损害过程中的作用。方法:40只ICR小鼠随机分为对照组和染毒组,每组20只,雌雄各半。经口灌胃给予小鼠50 mg/kg B(a)P,每周2次,持续8周,对照组同时给予等量的橄榄油,染毒结束后继续饲养8周。取肺脏组织,提取总RNA,采用SOliD高通量测序技术检测miRNAs表达谱,进行miRNAs差异表达分析,并用real time-PCR验证miRNAs表达,TargetScan,miRanda及picTar软件预测miRNAs可能调控的靶基因。结果:绝大部分miRNAs在肺组织的表达信号强度较低。与对照组相比,B(a)P染毒组小鼠肺组织中共有109个miRNAs发生差异表达,其中50个miRNAs表达上调,59个miRNAs表达下调。上调幅度最大的为7.43倍,下调幅度最大的为40.63倍。为验证测序的结果,取下调较为明显的miR-20b生行real time-PCR分析,结果显示与测序结果相一致,靶基因预测显示miR-20b可能调节与细胞增殖、周期、凋亡及肿瘤发生相关的蛋白。结论:长期高剂量B(a)P染毒可引起小鼠肺组织miRNAs表达产生特异性改变,差异表达miRNAs可能在B(a)P致机体健康损害过程中起着重要作用。     

Effect of high-dose benzo (a)pyrene on microRNAs expression in mice lung tissues

目的:研究长期高剂量苯并(a)芘[B(a)P]染毒对小鼠肺组织miRNAs表达谱的影响,探讨miRNAs在B(a)P健康损害过程中的作用.方法:40只1CR小鼠随机分为对照组和染毒组,每组20只,雌雄各半.经口灌胃给予小鼠50 mg/kg B (a)P,每周2次,持续8周,对照组同时给予等量的橄榄油,染毒结束后继续饲养8周.取肺脏组织,提取总RNA,采用SOliD高通量测序技术检测miRNAs表达谱,进行miRNAs差异表达分析,并用real time-PCR验证miRNAs表达,TargetScan,miRanda及picTar软件预测miRNAs可能调控的靶基因.结果:绝大部分miRNAs在肺组织的表达信号强度较低.与对照组相比,B(a)P染毒组小鼠肺组织中共有109个miRNAs发生差异表达,其中50个miRNAs表达上调,59个miRNAs表达下调.上调幅度最大的为7.43倍,下调幅度最大的为40.63倍.为验证测序的结果,取下调较为明显的miR-20b进行real time-PCR分析,结果显示与测序结果相一致,靶基因预测显示miR-20b可能调节与细胞增殖、周期、凋亡及肿瘤发生相关的蛋白.结论:长期高剂量B(a)P染毒可引起小鼠肺组织miRNAs表达产生特异性改变,差异表达miRNAs可能在B(a)P致机体健康损害过程中起着重要作用.     

MicroRNA在肺癌耐药中的作用

摘 要：微小RNA（microRNA,miRNA) 是一类短链、非编码小分子RNA,不同癌症治疗中特异性miRNAs失调参与了肿瘤细胞耐药的过程,进而调节肿瘤细胞对治疗的敏感性。研究表明miRNAs参与肺癌化疗及靶向的耐药过程。miRNAs具有克服肺癌耐药的潜能,同时能够增加肺癌细胞对化疗及靶向治疗的敏感性。全文综述了现阶段肺癌治疗中miRNAs在化疗药物及靶向药物耐药机制中的作用。     

外泌体在多发性骨髓瘤治疗中潜在作用

目的作为细胞间信息的重要传播者,外泌体在肿瘤诊断、治疗和预后等方面起着重要作用。研究证实其相关抑制剂或者类似物等具有抗多发性骨髓瘤(multiple myeloma,MM)的作用。本研究旨在总结国内外研究中外泌体在MM治疗中的潜在作用。方法以"外泌体、多发性骨髓瘤、治疗"为关键词,检索2004-08-2019-01PubMed和万方数据库检索系统中的文献。纳入标准:(1)外泌体的生物学功能;(2)外泌体在MM治疗中作用的研究。经筛选符合要求的文献有49篇。结果MM来源的外泌体某些成分如miRNA、蛋白质、lncRNA等与正常人之间表达存在差异,可利用其抑制剂或类似物发挥抗MM的作用,从而提供潜在治疗靶点。肿瘤来源外泌体及其加工修饰产物可作为一种新的癌症疫苗发挥抗肿瘤免疫;且外泌体的天然纳米结构使其成为药物递送的载体,可用于疾病的治疗。但目前疫苗相关研究较局限,药物载体在肿瘤治疗的应用方面研究亦甚少。结论外泌体可通过特异性表达某些成分而提供潜在治疗靶点,如其本身及其加工产物可制成疫苗,或以运载药物的形式在MM的治疗中发挥潜在作用。但目前这些研究尚处于初级阶段,外泌体应用于MM临床治疗尚需大量研究进一步证实。     

Hematological malignancies: role of miRNAs and their in silico aspects

Hematological malignancies is a broad term that includes blood cell cancers including chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), acute myeloid leukemia (AML), Myelodysplastic syndrome, acute lymphocytic leukemia (ALL), multiple myelomas (MM) and lymphomas. miRNAs are ~22-nt long non-coding RNAs that play a very important role in gene regulation by binding to mRNA at their complementary sequence. These miRNAs are conceptually connected with various signal and pathway networks that make them capable of regulating various diseases including hematological malignancies. These miRNAs are not only playing regulatory roles in hematological malignancies, but are also providing new potent markers for efficient diagnosis and prognosis for hematological malignancies patients. Since the discovery of very first miRNA, the importance and role of miRNAs have been established in various fields, and there is a need to search for new potent miRNAs and their targets. A large amount of sequence data have been generated in last few years, which has further generated the need to develop efficient and reliable computational tools to analyze and extract out relevant information promptly from raw data. Here, we review various possible roles played by miRNA in hematological malignancies, principles involved in miRNA gene identification, target prediction and their preceding role in hematological malignancies research.     

Genome-wide pharmacologic unmasking identifies tumor suppressive microRNAs in multiple myeloma

Epigenetic alterations have emerged as an important cause of microRNA (miRNA) deregulation. In Multiple Myeloma (MM), a few tumor suppressive miRNAs silenced by DNA hypermethylation have been reported, but so far there are few systemic investigations on epigenetically silenced miRNAs. We conducted genome-wide screening for tumor suppressive miRNAs epigenetically silenced in MM. Four Human MM Cell lines were treated with demethylating agent 5′azacytidine (5′aza). Consistently upregulated miRNAs include miR-155, miR-198, miR-135a*, miR-200c, miR-125a-3p, miR-188-5p, miR-483-5p, miR-663, and miR-630. Methylation array analysis revealed increased methylation at or near miRNA-associated CpG islands in MM patients. Ectopic restoration of miR-155, miR-198, miR-135a*, miR-200c, miR-663 and miR-483-5p significantly repressed MM cell proliferation, migration and colony formation. Furthermore, we derived a 33-gene signature from predicted miRNA target genes that were also upregulated in MM patients and associated with patient survival in three independent myeloma datasets. In summary, we have revealed important, epigenetically silenced tumor suppressive miRNAs by pharmacologic reversal of epigenetic silencing.     

DNA-demethylating and anti-tumor activity of synthetic miR-29b mimics in multiple myeloma

Aberrant DNA methylation plays a relevant role in multiple myeloma (MM) pathogenesis. MicroRNAs (miRNAs) are a class of small non-coding RNAs that recently emerged as master regulator of gene expression by targeting protein-coding mRNAs. However, miRNAs involvement in the regulation of the epigenetic machinery and their potential use as therapeutics in MM remain to be investigated. Here, we provide evidence that the expression of de novo DNA methyltransferases (DNMTs) is deregulated in MM cells. Moreover, we show that miR-29b targets DNMT3A and DNMT3B mRNAs and reduces global DNA methylation in MM cells. In vitro transfection of MM cells with synthetic miR-29b mimics significantly impairs cell cycle progression and also potentiates the growth-inhibitory effects induced by the demethylating agent 5-azacitidine. Most importantly, in vivo intratumor or systemic delivery of synthetic miR-29b mimics, in two clinically relevant murine models of human MM, including the SCID-synth-hu system, induces significant anti-tumor effects. All together, our findings demonstrate that aberrant DNMTs expression is efficiently modulated by tumor suppressive synthetic miR-29b mimics, indicating that methyloma modulation is a novel matter of investigation in miRNA-based therapy of MM.     

Potential relationship and clinical significance of miRNAs and Th17 cytokines in patients with multiple myeloma

We evaluated the potential relationship between miRNAs and Th17 cytokines in multiple myeloma (MM) patients. Twenty-seven newly diagnosed myeloma patients and eight normal donors were studied. We determined that the relative expression levels of miR-15a/16, miR-34a, miR-194 in MM patients were significantly lower than those in the healthy controls with exception for miR-181a/b, which showed significantly higher in MM patients (P < 0.05). In contrast, the levels of IL-17, IL-21 and IL-27 were up-regulated in MM patients compared to healthy controls while IL-22 was down-regulated (P < 0.05). The expression patterns of them were differentially present in various groups according to the International Staging System (ISS) criteria. Up-regulated IL-17, IL-21 and IL-27 may potentially down-regulate the expression of several miRNAs in MM patients. Establishment of the relationship may be useful for understanding the pathogenesis of MM and for clinical diagnosis of the disease.     

Downregulated plasma miR-92a levels have clinical impact on multiple myeloma and related disorders

Recent studies have demonstrated that one-third of known microRNAs (miRNAs) are stably detectable in plasma. Therefore, we assessed plasma miRNAs to investigate the dynamics of oncomir 17-92a, which is highly expressed in multiple myeloma (MM) patients. The plasma miR-92a level in symptomatic MM patients was significantly downregulated compared with normal subjects (P<0.0001), regardless of immunoglobulin subtypes or disease stage at diagnosis. In contrast, miR-92a levels in peripheral blood CD8(+) or CD4(+) cells from MM patients were lower than those of normal subjects, and the miR-92a levels of the cells tended to correlate with plasma miR-92a levels. The plasma miR-92a level in the complete remission group became normalized, whereas the partial response (PR) and very good PR groups did not reach the normal range. In smoldering MM, the plasma miR-92a level did not show a significant difference compared with normal subjects. Our findings suggest that measurement of the plasma miR-92a level in MM patients could be useful for initiation of chemotherapy and monitoring disease status, and the level may represent, in part, the T-cell immunity status of these patients.