长链非编码RNA DNAJC3-AS1、微RNA-214-3p在结肠癌组织、结肠腺瘤性息肉中的表达水平及临床意义

目的 探讨长链非编码RNA(lncRNA)DNAJC3-AS1、微RNA-214-3p(miR-214-3p)在结肠癌组织、结肠腺瘤性息肉中的表达水平及临床意义。方法 lnCAR数据库检索结肠癌组织、正常结肠组织中LncRNA DNAJC3-AS1表达情况;选取2016年5月至2019年1月北京市丰台中西医结合医院收治的86例结肠癌病人、53例结肠腺瘤性息肉病人为研究对象,收集结肠癌病人的结肠癌组织、对应癌旁组织及结肠腺瘤性息肉病人的结肠腺瘤性息肉,实时荧光定量PCR(qRT-PCR)法测定组织中lncRNA DNAJC3-AS1、miR-214-3p表达水平;分析结肠癌组织lncRNA DNAJC3-AS1、miR-214-3p表达水平与结肠癌病人临床病理特征、预后关系;分析结肠癌组织中lncRNA DNAJC3-AS1表达水平与miR-214-3p相关性,生物信息学预测lncRNA DNAJC3-AS1与miR-214-3p的关系。结果 lnCAR数据库分析显示,结肠癌组织lncRNA DNAJC3-AS1表达水平（4.40±0.49）高于正常结肠组织（4.00±0.36）(P&l...     

血清长基因间非编码RNA00511、微RNA-515-5p在子宫内膜癌中表达与临床病理特征及预后的关系

目的 探究子宫内膜癌病人血清长基因间非编码RNA00511(LINC00511)、微RNA-515-5p(miR-515-5p)表达水平与临床病理特征及预后的关系。方法 收集2017年3月至2019年2月海南医学院第二附属医院子宫内膜癌病人（子宫内膜癌组）82例;同时收集子宫内膜不典型增生病人（不典型增生组）67例及健康体检者（对照组）79例。采用实时荧光定量逆转录聚合酶链式反应（qRT-PCR）检测血清LINC00511、miR-515-5p表达水平;采用Pearson相关分析子宫内膜癌病人血清LINC00511与miR-515-5p表达水平的相关性;采用Kaplan-Meier法分析子宫内膜癌病人血清LINC00511、miR-515-5p表达水平与预后的关系;多因素Cox回归分析影响子宫内膜癌病人预后的因素。结果 对照组、不典型增生组、子宫内膜癌组血清LINC00511表达水平（1.02±0.21、1.82±0.31、2.60±0.45）依次升高,miR-515-5p表达水平（1.01±0.20、0.70±0.18、0.42±0.16）依次降低（P<0.05）。肌层浸润深度...     

金丝桃苷抑制鼻咽癌细胞生物学行为的分子机制研究

目的 探讨金丝桃苷对鼻咽癌细胞增殖、迁移和侵袭等生物学行为的影响和机制。方法 采用10、20及40 mg/L金丝桃苷的培养液孵育SUNE1细胞，依次记为低、中、高剂量组；将长链非编码RNA(lncRNA)配对盒基因8反义RNA 1(PAX8-AS1)过表达或敲低质粒、miR-494-3p模拟物及其阴性对照分别转染至40 mg/L金丝桃苷处理的SUNE1细胞，采用CCK-8实验、平板克隆实验、Transwell实验检测SUNE1细胞活力、克隆形成以及迁移和侵袭。蛋白质印记法分析基质金属蛋白酶（MMP）-2和MMP-9蛋白表达。实时荧光定量聚合酶链反应（RT-qPCR）检测lncRNA PAX8-AS1和miR-494-3p表达。双荧光素酶报告实验分析lncRNA PAX8-AS1和miR-494-3p靶向关系。结果低、中、高剂量金丝桃苷降低SUNE1细胞活力、迁移数和侵袭数，下调MMP-2蛋白、MMP-9蛋白以及miR-494-3p表达水平，上调lncRNA PAX8-AS1表达水平（P<0.05）。lncRNA PAX8-AS1靶向负调控miR-494-3p表达。过表达lncRN...     

结合外周血微RNA-181b表达水平探讨急性脑梗死病人侧支循环形成的影响因素

目的 结合外周血微RNA(miR)-181b表达水平探讨急性脑梗死病人侧支循环形成的影响因素。方法 选择2015年6月至2019年5月在深圳市龙华区中心医院住院治疗的ACI病人175例,采用计算机产生随机数按3∶1的比例将纳入病人分为训练集（129例）和测试集（46例）,其中129例训练集病人根据侧支循环形成情况分为两组,其中侧支循环良好组病人90例,侧支循环不良组病人39例,采用定量逆转录PCR(RT-qPCR)检测血清miR-181b表达水平,采用Spearman相关法分析ACI病人血清miR-181b表达与侧支循环形成的相关性,采用logistic回归法分析ACI病人侧支循环形成不良的影响因素,并建立预测模型。结果 侧支循环良好组病人血清miR-181b表达1.92±0.19显著高于侧支循环形成不良组1.48±0.26(P<0.05),Spearman相关性分析结果表明,血清miR-181b表达与侧支循环形成呈显著正相关关系（r=0.77,P<0.001）。logistic回归分析结果表明高血压病史高血压史、血同型半胱氨酸（Hcy）、后循环病变均为ACI病人侧支循环形...     

长链非编码小核仁 RNA宿主基因 7调控微小 RNA-331-3p表达影响甲状腺癌细胞增殖、迁移和侵袭的机制研究

目的 探讨长链非编码小核仁RNA宿主基因7(lncRNA SNHG7)对甲状腺癌细胞增殖、迁移和侵袭的影响以及潜在的作用机制.方法 本研究起止时间为2018年1月至2019年2月,人甲状腺细胞Nthy-ori 3-1和人甲状腺癌细胞K1、BCPAP、TPC-1购自中国科学院上海细胞库,用qRT-PCR检测SNHG7和微小RNA-331-3p(miR-331-3p)的表达水平;将si-NC组(转染SNHG7干扰表达载体阴性对照)、si-SNHG7组(转染SNHG7干扰表达载体)、si-SNHG7+anti-miR-NC组(共转染SNHG7干扰表达载体和miR-331-3p抑制剂阴性对照)、si-SNHG7+anti-miR-331-3p组(共转染SNHG7干扰表达载体和miR-331-3p抑制剂),均用脂质体法转染至TPC-1细胞;采用蛋白质印迹法(Western Blotting)检测蛋白表达;四甲基偶氮唑盐比色法(MTT)检测细胞增殖;Transwell检测细胞迁移和侵袭;双荧光素酶报告基因检测实验检测荧光活性.结果 与人甲状腺细胞Nthy-ori 3-1相比,人甲状腺癌细胞K1、BCPAP、TPC-1中SNHG7的表达水平显著升高[(1.42±0.16)、(1.51±0.18)、(2.56±0.15)比(1.01±0.05)];miR-331-3p的表达水平显著下降[(0.63±0.11)、(0.60±0.10)、(0.42±0.08)比(1.00±0.06)],差异有统计学意义(P<0.05).与si-NC组相比,si-SNHG7组TPC-1细胞中细胞周期蛋白D1(Cyclin D1)、神经钙黏蛋白(N-cadherin)的表达水平显著降低,细胞周期蛋白依赖性激酶抑制剂1A(p21)、上皮钙黏蛋白(E-cadherin)的表达水平显著升高,TPC-1细胞活性显著降低[24 h:(0.20±0.06)比(0.49±0.10),48 h:(0.50±0.13)比(1.10±0.22),72 h:(0.60±0.13)比(1.60±0.10)],迁移细胞数降低[(63±8.97)个比(144±11.36)个],侵袭细胞数降低[(55±10.03)个比(136±13.38)个],差异有统计学意义(P<0.05).SNHG7可靶向调控miR-331-3p的表达.抑制miR-331-3p表达可逆转干扰SNHG7对TPC-1细胞的作用.SNHG7可靶向调控miR-331-3p的表达.抑制miR-331-3p表达可逆转干扰SNHG7对TPC-1细胞的作用.结论 干扰SNHG7可通过上调miR-331-3p抑制TPC-1细胞的恶性生物学行为.     

长链非编码RNA OPA相互作用蛋白5反向转录序列1靶向调控微RNA-128-3p对结直肠癌细胞增殖、侵袭和转移的影响

目的 探究长链非编码RNA OPA相互作用蛋白5反向转录序列1(lncRNA OIP5-AS1)在结直肠癌中的表达情况及靶向调控微RNA-128-3p(miR-128-3p)对结直肠癌细胞增殖、侵袭和转移的影响。方法 采用实时荧光定量PCR(qPCR)定量分析2017年1月至2018年12月在郑州大学人民医院住院并进行手术治疗的38例结直肠癌病人癌组织及相应癌旁组织、结直肠癌细胞系（SW480、SW620、HT-29和LoVo）及人正常结直肠黏膜细胞FHC中lncRNA OIP5-AS1和miR-128-3p的表达。将SW620细胞设为si-OIP5-AS1组、si-NC组、miR-128-3p mimic组、mimic-NC组、miR-128-3p inhibitor+si-OIP5-AS1组和inhibitor-NC+siOIP5-AS1组,采用细胞计数试剂盒（CCK-8）实验和克隆形成实验检测细胞增殖能力,采用划痕实验和transwell实验检测细胞侵袭与迁移能力,采用蛋白质印迹法检测E2F1、细胞周期蛋白D1(Cyclin D1)、波形蛋白（Vimentin）、N-钙黏蛋白（N...     

长链非编码 RNAMIR22HG和微 RNA-9-3p在骨关节炎病人血清中的表达及与病情严重程度的相关性

目的探究长链非编码 RNA(lncRNA)MIR22HG和微 RNA-9-3p(miR-9-3p)在骨关节炎( OA)病人血清中的表达及与病情严重程度的相关性。方法选取 2020年 10月至 2021年 10月在南充市中医医院确诊为 OA的病人 120例,作为 OA组, 60例体检健康人群作为对照组;采用实时荧光定量逆转录聚合酶链式反应( qRT-PCR)检测血清 lncRNA MIR22HG和 miR-9-3p的表达水平; Pearson法分析 lncRNA MIR22HG和 miR-9-3p的相关性以及二者与视觉模拟( VAS)评分、美国特种外科医院膝关节评分( HSS)以及西安大略和麦马斯特大学骨关节炎指数可视化量表( WOMAC)评分的相关性;绘制受试者操作特征( ROC)曲线分析 lncRNA MIR22HG和 miR-9-3p对 OA的诊断价值。结果 OA组的血清中 lncRNA MIR22HG表达水平显著高于对照组(1.48±0.25比 1.01±0.22)miR-9-3p的表达水平显著低于对照组( 0.72±0.13比 1.05±0.12)(P<0.05);相关性分析显示, lncRNA MIR22HG和 miR-9-3p的表,达水平呈负相关关系( P<0.05); KL Ⅳ级血清 lncRNA MIR22HG的表达水平、 VAS评分、 WOMAC评分显著高于 KL Ⅲ级和 KL Ⅱ级, KL Ⅲ级显著高于 KL Ⅱ级( P<0.05),KL Ⅳ级血清 miR-9-3p的表达水平、 HSS评分显著低于 KL Ⅲ级和 KL Ⅱ级, KL Ⅲ级显著低于 KL Ⅱ级( P<0.05); OA病人 lncRNA MIR22HG的表达水平与 VAS评分、 WOMAC评分均呈正相关,与 HSS评分呈负相关( P<0.05); miR-9-3p表达水平与 VAS评分、 WOMAC评分均呈负相关,与 HSS评分呈正相关( P<0.05);二者联合诊断的 AUC高于 lncRNA MIR22HG、miR-9-3p单独诊断的 AUC值( Z=2.22,P=0.012;Z=1.43,P=0.025)。结论 OA病人血清中 lncRNA MIR22HG表达水平显著升高, miR-9-3p表达水平显著降低,二者与 OA病情严重程度密切相关,且对 OA具有一定的诊断价值。     

lncRNA FOXD2-AS1靶向miR-3194-3p对口腔鳞癌细胞增殖及凋亡影响的体外研究

摘要：目的:探讨lncRNA FOXD2-AS1对口腔鳞癌细胞增殖及凋亡的影响及分子机制。方法:实时荧光定量PCR（RT-qPCR）检测口腔鳞癌细胞系中lncRNA FOXD2-AS1和miR-3194-3p的表达水平;将FOXD2-AS1干扰表达载体、miR-3194-3p模拟物、miR-3194-3p抑制剂+FOXD2-AS1干扰表达载体转染至CAL27细胞中,Western blot法检测蛋白表达; MTT检测细胞存活率;流式细胞术检测细胞凋亡;双荧光素酶报告实验检测lncRNA FOXD2-AS1和miR-3194-3p的靶向关系。结果:口腔鳞癌细胞系中lncRNA FOXD2-AS1表达水平升高,miR-3194-3p表达水平降低（P<0.01）。lncRNA FOXD2-AS1低表达或miR-3194-3p高表达,口腔鳞癌CAL27细胞中细胞周期蛋白D1（Cyclin D1）表达水平和细胞存活率降低,裂解的半胱氨酸天冬氨酸蛋白酶-3（cleaved-caspase-3）表达水平和细胞凋亡率升高（P<0.01）;且lncRNA FOXD2-AS1低表达降低了β-连环蛋白（β-catenin）蛋白表达。lncRNA FOXD2-AS1靶向调控miR-3194-3p,低表达miR-3194-3p逆转了FOXD2-AS1低表达对口腔鳞癌细胞CAL27增殖和凋亡及Wnt/β-catenin信号通路的影响。结论:下调lncRNA FOXD2-AS1表达可能通过上调miR-3194-3p抑制Wnt/β-catenin信号通路抑制口腔鳞癌细胞增殖,促进细胞凋亡。     

长链非编码RNA SNHG3调节微小RNA-340-5p/Skp2轴对胃癌细胞恶性生物学行为的影响研究

目的 探讨长链非编码(long non-coding,Lnc)RNA SNHG3对胃癌细胞恶性生物学行为的影响及其分子机制。方法 实时定量聚合酶链式反应法检测人正常胃黏膜上皮细胞及胃癌细胞中LncRNA SNHG3表达水平。分别构建SNHG3敲低表达组和阴性对照组,并设置空白对照组。采用细胞计数-8法,Transwell实验及流式细胞技术检测敲低LncRNA SNHG3表达对胃癌细胞增殖、迁移、侵袭及凋亡的影响。通过生物学信息网站预测LncRNA SNHG3下游靶基因,利用双荧光素酶报告实验验证其结合关系。蛋白质印迹法检测LncRNA SNHG3对下游靶蛋白表达的影响。结果 胃癌细胞HGC-27、MGC-803、MKN-45及BGC-823中LncRNA SNHG3表达水平明显高于正常胃黏膜上皮细胞CES-1,差异有统计学意义(F=16.128,P <0.05)。敲低SNHG3表达后胃癌细胞增殖、迁移及侵袭能力明显降低,细胞凋亡比例明显升高。LncRNA SNHG3、Skp2均与微小RNA(micro RNA,miR)-340-5p之间存在结合位点,且miR-340-5p可降低W...     

慢性乙型病毒性肝炎病人血浆中LncRNA PVT1、微RNA-31-5p表达水平与炎症损伤程度的相关性分析

目的 探讨长链非编码RNA(LncRNA)人浆细胞瘤转化迁移基因1（）、微RNA-31-5p(miR-31-5p)在慢性乙型病毒性肝炎（CHB）病人血浆中的表达情况,并分析其与炎症损伤程度的关系。方法 选取东莞市人民医院2019年12月至2020年12月确诊的180例CHB病人为CHB组,所有病人均行肝穿刺活检,根据CHB病人炎症损伤程度分为G1组（66例）,G2组（58例）,G3组（41例）,G4组（15例）,根据肝纤维化程度进行分组,无明显肝纤维化（S0,S1）组（64例）,明显肝纤维化组（S2,S3）(78例),早期肝硬化（S4）组（38例）。同时选取55例健康者为对照组。收集CHB组和对照组的一般资料,分别检测两组血清肿瘤坏死因子-α(TNF-α)、白细胞介素（IL）-6、IL-1β、IL-10、超敏C反应蛋白（Hs-CRP）、丙氨酸氨基转移酶（ALT）、天冬氨酸转氨酶（AST）、谷氨酰基转移酶（GGT）等指标水平;采用实时荧光定量PCR法对血清中LncRNA PVT1、miR-31-5p表达水平进行检测,Pearson法分析LncRNA PVT1、miR-31-5p表达的相关...     

RNA therapeutics: beyond RNA interference and antisense oligonucleotides

Here, we discuss three RNA-based therapeutic technologies exploiting various oligonucleotides that bind to RNA by base pairing in a sequence-specific manner yet have different mechanisms of action and effects. RNA interference and antisense oligonucleotides downregulate gene expression by inducing enzyme-dependent degradation of targeted mRNA. Steric-blocking oligonucleotides block the access of cellular machinery to pre-mRNA and mRNA without degrading the RNA. Through this mechanism, steric-blocking oligonucleotides can redirect alternative splicing, repair defective RNA, restore protein production or downregulate gene expression. Moreover, they can be extensively chemically modified to acquire more drug-like properties. The ability of RNA-blocking oligonucleotides to restore gene function makes them best suited for the treatment of genetic disorders. Positive results from clinical trials for the treatment of Duchenne muscular dystrophy show that this technology is close to achieving its clinical potential.     

血小板RNA与肿瘤

血小板是人体中重要的无核血细胞,生理状态下主要发挥止血和促进伤口愈合的作用。此外,血小板也参与了多种病理过程,主要包括炎症、血管生成、组织再生以及肿瘤的恶性发展等。目前,大量研究显示血小板在肿瘤发生发展中发挥着极其重要的作用,并且参与了肿瘤发生发展的多个过程。最新的研究证明肿瘤微环境中血小板RNA水平发生明显改变并且影响着肿瘤的发展进程。该文综述了近几年报道的在肿瘤微环境中肿瘤细胞对血小板RNA影响及其可能产生的病理效应的相关研究。为临床肿瘤检测以及基于RNA进行血小板与肿瘤之间交互作用的研究提供一定指导。     

Effects of 5-fluorocytidine on mammalian transfer RNA and transfer RNA methyltransferases.

Effects of 5-fluorocytidine on mouse liver tRNA and mouse liver tRNA methyltransferases were investigated. tRNA isolated from drug-treated tissue was shown to contain 5-fluorocytidine and 5 fluorouridine. The amounts of all 5-substituted pyrimidine nucleosides such as 5-methylcytidine, 5-methyluridine, pseudouridine and 5,6-dihydrouridine were substantially reduced. The decreased methylation of tRNA was shown to result from decreased tRNA cytosine-5-methyltransferase and tRNA uracil-5-methyltransferase activities and capacities. Incorporation of 5-fluorouridine into tRNA, as well as the effects of 5-fluorocytidine administration on the modified uridine derivatives in tRNA, suggested the in vivo conversion of 5-fluorocytosine derivatives to 5-fluorouracil derivatives, as administration of the latter had been shown previously to cause the same effects. The inhibition of tRNA cytosine-5-methyltransferase after administration of 5-fluorocytidine resembles the effect of another cytidine analog, 5-azacytidine, which is known to cause lack of 5-methylcytidine in mouse liver tRNA and the inhibition of this particular tRNA methyltransferase.     

Cyclophilin B stimulates RNA synthesis by the HCV RNA dependent RNA polymerase

Cyclophilins are cellular peptidyl isomerases that have been implicated in regulating hepatitis C virus (HCV) replication. Cyclophilin B (CypB) is a target of cyclosporin A (CsA), an immunosuppressive drug recently shown to suppress HCV replication in cell culture. Watashi et al. recently demonstrated that CypB is important for efficient HCV replication, and proposed that it mediates the anti-HCV effects of CsA through an interaction with NS5B [Watashi K, Ishii N, Hijikata M, Inoue D, Murata T, Miyanari Y, et al. Cyclophilin B is a functional regulator of hepatitis C virus RNA polymerase. Mol Cell 2005;19:111-22]. We examined the effects of purified CypB proteins on the enzymatic activity of NS5B. Recombinant CypB purified from insect cells directly stimulated NS5B-catalyzed RNA synthesis. CypB increased RNA synthesis by NS5B derived from genotype 1a, 1b, and 2a HCV strains. Stimulation appears to arise from an increase in productive RNA binding. NS5B residue Pro540, a previously proposed target of CypB peptidyl-prolyl isomerase activity, is not required for stimulation of RNA synthesis.     

RNA interference and potential applications

RNA interference (RNAi) is the process of using specific sequences of double-stranded RNA (dsRNA) to knock down the expression level of sequence-homologous genes. Such ability of small interfering RNA (siRNA) in mammalian cells will undoubtedly revolutionize the study of functional genomics, the discovery of drug targets and even the treatment of human diseases. In this review we briefly describe the history of RNAi discovery, the RNAi mechanism and the general guideline for siRNA design as well as various methods for siRNA production and delivery. We also introduce the potential applications of siRNA, inducible siRNA and siRNA library in speeding up basic biomedical research and in acting as potential therapeutic agents for treatment of numerous human diseases.     

RNA interference and innate immunity

RNA interference is an evolutionarily conserved gene silencing process triggered by double-stranded RNAs. Common to all cell types, is the production of 21-24 nucleotide small interfering RNA (siRNAs), which guide the RNA-induced silencing complex (RISC) to identify and cleave target mRNA sequences. Presently, this biological breakthrough method has revolutionised gene function studies and holds great promise as validating drug targets and treating human diseases. However, despite the success that has been achieved by this technology, studies carried in human blood cells have revealed that siRNAs could generate bystander effects, including the activation of innate immunity and inhibition of unintended target genes. Interestingly, 2' uridine-modified siRNAs did not trigger TLR signalling, but they totally suppressed immune activation by immunostimulatory siRNAs when both molecules where delivered to the same endosomes. This review describes the recent advances in understanding the innate immune response to both single and double-stranded siRNAs. Also, it highlights the spectrum of molecular strategies allowing the design of therapeutic siRNAs with minimal side effects.     

Oxidative RNA damage and neurodegeneration

Although cellular RNA should be subject to the same oxidative insults as DNA and other cellular macromolecules, oxidative damage to RNA has not been a major focus in investigating the magnitude and the biological consequences of the free radical damage. However, because RNA is mostly single-stranded and its bases are not protected by hydrogen bonding and are less protected by specific proteins, RNA may be more susceptible to oxidative insults than DNA. Thereafter, oxidative damage to protein-coding RNA or non-coding RNA will potentially cause errors in proteins or dysregulation of gene expression. While less lethal than mutations in genome, such non-acutely lethal insults to cells might be associated with underlying mechanisms of several human diseases, especially chronic degeneration. Recently, oxidative RNA damage has been described in several neurodegenerative diseases including Alzheimer disease, Parkinson disease, dementia with Lewy bodies, and prion diseases. Of particular interest, oxidative RNA damage is a feature in vulnerable neurons at the very earliest-stages of these diseases, suggesting that RNA oxidation may actively contribute to the onset or to the development of disease. Mechanistically speaking, an increasing body of evidence suggests that the detrimental effects of oxidative RNA damage to protein synthesis are attenuated, at least in part, by the existence of mechanisms that avoid the incorporation of the damaged ribonucleotides into the translational machinery. Further investigations toward understanding of the consequences and processing mechanisms related to oxidative RNA damage may provide significant insights into the pathogenesis and therapeutic strategies for neurodegenerative and other degenerative diseases.     

RNA Interference and Cancer Therapy

Since its discovery in 1998, RNA interference (RNAi) has revolutionized basic and clinical research. Small RNAs, including small interfering RNA (siRNA), short hairpin RNA (shRNA) and microRNA (miRNA), mediate RNAi effects through either cleavage-dependent or cleavage-independent RNA inducible silencing complex (RISC) effector processes. As a result of its efficacy and potential, RNAi has been elevated to the status of “blockbuster therapeutic” alongside recombinant protein and monoclonal antibody. RNAi has already contributed to our understanding of neoplasia and has great promise for anti-cancer therapeutics, particularly so for personalized cancer therapy. Despite this potential, several hurdles have to be overcome for successful development of RNAi-based pharmaceuticals. This review will discuss the potential for, challenges to, and the current status of RNAi-based cancer therapeutics.