猪miR-378种子序列的多态性对其功能以及胴体性状的影响

本研究旨在探索miR-378种子序列多态性对其结构、功能和猪胴体性状的影响.利用荣昌猪和亚洲野猪的单核苷酸多态性(SNP)进行群体遗传选择分析,预测miR-378种子序列的多态性对其结构的改变,统计不同猪种中等位基因频率;采用microRNA类似物验证种子序列多态性对miR-378在脂肪细胞中功能以及相关基因表达的影响...     

miR-378促进脂质生成相关靶基因鉴定

旨为验证miR-378在脂肪细胞中的功能,及其脂质相关靶基因的筛选和鉴定.利用miR-378类似物转染3T3-L1细胞,验证miR-378在脂肪细胞中的功能;根据靶标位点的保守性以及促脂功能确定miR-378潜在靶基因;采用microRNA pulldown技术验证miR-378与靶基因的靶标关系;运用双荧光素报告基因...     

牦牛miR-378前体克隆及组织表达分析

旨在克隆牦牛miR-378的前体序列,阐明其组织表达规律,结合bta-miR-378靶基因的生物信息学预测和分析,探讨miR-378在牦牛生长发育过程中的调控功能。采用PCR方法成功克隆类乌齐牦牛miR-378前体序列,实时荧光定量PCR(RT-qPCR)检测miR-378-3p在各组织中的表达模式,结合生物信息学软件TargetScan、DAVID以及数据库NCBI、miRbase等对miR-378进行保守性分析、靶基因预测及其生物学功能分析。结果表明,miR-378在各物种间高度保守,且miR-378-3p在各组织中广泛表达,其中在臀大肌中表达水平最高,显著高于其他组织(P<0.01),在臀脂中的表达高于卵巢、大脑、乳腺和肝脏。获得的272个靶基因主要参与细胞分化、细胞发育、大分子代谢等多个生物学过程,涉及孕酮介导的卵母细胞成熟、促性腺激素释放激素(GnRH)信号通路等,由此推测,miR-378可能在卵泡发育、卵母细胞成熟过程中起关键作用,进而影响母牦牛的繁殖性能。     

Mir-378a-5p抑制鼻咽癌细胞系CNE-1 细胞增殖以及肿瘤生长的初步研究*

目的： MiR-378a-5p是一种被认为在多种肿瘤发生过程中具有抑制肿瘤生长的微小RNA。然而miR-378a-5p在鼻咽癌中的 作用尚未见报道。因此,本文旨在通过临床样本的miRNA 表达谱分析以及细胞学实验从而揭示miR-378a-5p在鼻咽癌肿瘤发生过程中的作用。方法与结果：我们通过生物信息学的方法获取了鼻咽癌临床样本中miR-378a-5p的表达信息并通过与正常组织的 对比发现miR-378a-5p在鼻咽癌肿瘤组织中表达水平显著降低（P<0.01）。其次,我们发现高表达miR-378a-5p的鼻咽癌CNE-1 细 胞增殖速度显著较对照组降低（约40%~50%）。克隆形成实验证实了瞬时转染miR-378a-5p的鼻咽癌CNE-1 细胞的克隆形成数 量显著减弱。我们通过将稳定表达miR-378a-5p的CNE-1 细胞注射到裸鼠体内形成移植瘤并记录肿瘤生长曲线,结果显示 miR-378a-5p高表达组的裸鼠移植瘤体积明显较对照组小约50%,肿瘤重量显著降低(对照组0.33 g,处理组0.15 g)。结论：本研究通过对临床样本的分析以及在细胞和动物水平的实验验证揭示了miR-378a-5p具有抑制鼻咽癌肿瘤细胞增殖和肿瘤生长的作用。     

采用双荧光素酶报告法验证乳腺癌中miR-155靶标

MiR-155与乳腺癌发生发展密切相关. 本研究以乳腺癌组织和血清中均显著高表达的miR 155为研究对象,利用TargetScan和PicTar预测miR 155的靶基因. 根据miR 155与靶基因结合的保守性、动力学及靶基因功能,筛选出miR 155的靶基因ACTA1（actin alpha 1, skeletal muscle）和CEBPB（CCAAT/enhancer binding protein beta）,并用双荧光素酶报告法验证. 将ACTA1和CEBPB的3′-UTR（3′-untranslated region）全长序列载入海肾荧光素酶基因的下游,并构建结合位点的突变序列,得到pRL-TK-Aw、pRL-TK-Am、pRL-TK-Cw、pRL-TK-Cm载体.不同海肾荧光素酶载体转染Bcap37乳腺癌细胞,同时转染miR-155及内参对照萤火虫荧光素酶载体pGL3-control. 根据不同转染的海肾荧光素酶表达活性,运用SPSS软件分析,结果显示,CEBPB是miR-155在乳腺癌中的直接靶基因（P< 0.05）. miR-155通过下调CEBPB影响乳腺癌的发生.     

miR-26家族与肿瘤关系的研究进展

miR-26家族是由miR-26a、miR-26b、miR-1297及miR-4465等序列相似、结构相仿、种子区序列相同(UCAAGUA)的微小RNA(microRNAs,miRNAs)组成。多种组织细胞分化过程中伴有mi R-26家族表达的增加,其异常表达与特发性肺纤维化、原发性胆汁性肝硬化、多发性硬化及阿尔茨海默病等有关。近年研究报道,多种肿瘤组织亦存在mi R-26基因表达紊乱,并与肿瘤的发生发展密切相关。就miR-26家族及其调控的靶基因与肿瘤关系的研究进展进行综述。     

汉滩病毒M片段的核苷酸序列变异和系统发生树分析

The total RNA was extracted from two clinical blood samples of HFRS patients and the RNA was amplified by RT-PCRThe amplified DNA fragment of sample 613 involved nucleotides 1,471-1,873,and the sample 226 involved 540-1,244 nucleotides of M fragment of Hantaan virusThen the amplified PCR products were sequenced directlyThe sequencing results demonstrated that there was 84% identity between sample 613 and HV114 virus strain,but was 99% between sample 613 and HTN76-118 strainHowever,in sample 226,there was 95% sequence identity with HV114,82% with HTN76-118The results of phylogenetic tree analysis showed that HV613 was located in the same linage with HTN76-118,the HV226 was in the same linage with HV114 and A9 strains     

miR-92a家族与肿瘤关系的研究进展

miR-92a家族基因是由miR-25、miR-92a~1、miR-92a~2和miR-363等序列相似、结构相仿、种子区序列相同的微小RNA(microRNAs)组成,它们分别来自在进化过程中高度保守并互为旁系同源序列的miR-106b~25、miR-17~92和miR-106a~363基因簇。目前研究认为,miR-92a家族基因是一组与血管内皮细胞形成有关的miRNAs,其表达紊乱与肿瘤的发生发展密切相关。就miR-92a家族基因及其靶基因与肿瘤关系的研究进展进行综述。     

细胞质雄性不育高粱叶绿体 ndh D 基因的序列变异

片段SAAU－02 700特异地扩增自7种具可育细胞质的高粱材料的总DNA,含有叶绿体psa C(88bp)和ndh D(192bp)基因的部分序列。该片段与Eco Ri HindⅢ酶切的总DNA,线粒体DNA和叶绿体DNA杂交,在总DNA中获得了0.74kb的杂交带,而在叶绿体中获得0.74kb和0．45kb两条杂交带。与线粒体DNA无杂交;与经Hae Ⅲ酶切的总DNA杂交,在不育系中获得4．9kb的杂交带,而保持系的杂交带为4．45kb。参考GenBank中高粱的近缘物种玉米叶绿体基因组的序列,构建了ndh D基因区的酶切位点图谱,借此分析得出高粱不育系的叶绿体ndh D基因序列已发生改变。这种变异与高粱细胞质雄性不育反生的关系正在探讨中。     

生物序列的聚类分析

通过不同的聚类方式,对公共数据库中生物序列数据进行生物信息的挖掘,以达到在更广泛和更深入的框架中了解它们之间的相互关系的目的。以帕金森病相关基因所对应的mRNA序列为例,使用双序列比对的得分值作为序列之间的距离定义。同时为解决不同聚类分析之间的差异,分别采用模糊聚类和层次聚类两种不同的方法进行聚类分析。并由不同聚类方法得到的一致分类聚类的结果为基因功能分类提供支持,为进一步揭示生物序列所蕴涵的生物学知识和生物学规律提供可参考的依据。     

Identification of serum miR-378 and miR-575 as diagnostic indicators and predicting surgical prognosis in human epilepsy

BackgroundEpilepsy (EP) is a common neurological disorder which is characterized by excessive abnormal synchronization of neuronal discharges in the brain due to chronic recurrent seizures of multiple etiologies. Variety of microRNAs have been associated with the occurrence and development of EP. This study aimed to determine the aberrant expression of miR-378 and miR-575 in EP patients to validate their potential to distinguish EP from healthy patients.MethodsRT-qPCR was used to determine the expressions of miR-378 and miR-575 from serum specimens of 106 EP and 103 control individuals. Clinical indicators between EP patients and controls were assessed. Based on surgical outcome, EP patients were further divided into Engel I-IV EP. The potentials of miR-378 and miR-575 in discriminating EP from healthy participants and predicting surgical prognosis were calculated by receiver operating characteristic (ROC) analysis.ResultsWe found the miR-378 and miR-575 were significantly declined (P<0.001) in Engel I-II and III-IV EP patients with no difference in clinical parameters compared. Moreover, miR-378 and miR-575 displayed high sensitivity, specificity, and accuracy in distinguishing EP patients and predicting surgical outcomes. Moreover, after surgical treatment, miR-378 and miR-575 levels were increased compared with those at admission, suggesting their potentials in treatment response.ConclusionsmiR-378 and miR-575 could be utilized as novel and non-invasive serum biomarkers in discriminating EP from healthy controls and predicting surgical outcome, shedding new insights on epileptogenesis and EP treatment.     

microRNA-378 与肿瘤血管生成的研究进展

微小RNA(MicroRNAs,mi RNAs)是真核生物中一类长度约为21到23个核苷酸的非编码小分子单链RNA。mi RNA通过与靶m RNA 3′UTR(3′-untranslated region,3′非编码区)完全或不完全结合,抑制翻译或直接诱导其降解,发挥转录后负调控作用。mi RNA参与机体多种生理和病理过程,且可通过调控其靶标基因参与各种信号通路,影响血管生成。mi R-378属于诸多mi RNAs中的一种。目前已知mi R-378的研究主要集中在肿瘤发生及血管生成、心血管疾病和脑缺血等病理过程,其中与肿瘤发生及血管生成相关研究居多。mi R-378在不同肿瘤中的发挥的作用也不一样,在脑胶质瘤,肺癌,横纹肌肉瘤等肿瘤中发挥促癌基因的作用,在卵巢癌,胃癌,大肠癌等肿瘤中发挥抑癌基因的作用。但是,mi R-378调节肿瘤血管生成的作用机制还有待于深入研究。本文主要对mi R-378在四种肿瘤(脑胶质瘤、肺腺癌、卵巢癌和横纹肌肉瘤)中调控血管生成的相关性研究进展进行综述,以期为这些疾病的治疗和预防提供一种新的思路。     

急性冠脉综合征患者外周血miR-378a-3p的表达及其诊断价值

摘要 目的：探讨急性冠脉综合征（ACS）患者外周血miR-378a-3p的表达及其诊断价值。方法：收集ACS患者和健康人（control）血清各3份,进行基因芯片检测筛选发现miR-378a-3p差异表达最为显著。利用网络药理学分析miR-378a-3p与心肌缺血的相关性。构建小鼠心肌缺血再灌注（IR）模型,按缺血再灌注时间不同分为假手术（sham）组、I/R 1h组、I/R 3h组、I/R 6h组、I/R 12h组,检测外周血miR-378a-3p表达水平以及心肌肌钙蛋白T（cTnT）浓度水平。收集ACS患者（101例）和同期健康体检人群（49例）,进行实时定量PCR,检测miR-378a-3p在ACS患者的表达。采用受试者工作特征曲线（ROC）分析miR-378a-3p诊断ACS的诊断效能,并对ACS进行风险评估。结果：基因芯片结果显示miR-378a-3p在ACS患者外周血中表达升高。网络药理学分析提示,miR-378a-3p与心肌缺血存在共同靶点且miR-378a-3p可能通过细胞凋亡和血管生成途径干预心肌缺血,有作为心肌缺血标志物的可能。动物实验证实miR-378a-3p表达量随心肌缺血时间变化逐渐上调且在6h时表达明显升高,且miR-378a-3p表达水平较cTnT更早在心肌缺血中出现高表达。临床数据分析表明,同健康对照组比较,ACS患者血清中的miR-378a-3p在发生ACS的临床症状后,在短时间内可以显著升高。miR-378a-3p作为ACS的特异性诊断指标时,曲线下面积（AUC）为0.8476（95%可信区间为0.7749- 0.9204, P<0.001）,约登指数为0.6981,采用0.214作为cut-off值,miR-378a-3p诊断ACS的灵敏度和特异度分别为86.14%和83.76%。进一步的研究显示,在缺血的转化过程中,根据疾病分类的不同,结果显示miR-378a-3p的表达趋势也不同,在不稳定型心绞痛（UA）和心肌梗死（MI）组中,miR-378a-3p都显著上调（P<0.001）,同MI患者相比,UA患者miR-378a-3p表达上调更显著（P<0.05）,提示在诊断和判断ACS的分类中有较好的价值。结论：ACS患者外周血中miR-378a-3p表达升高是ACS发生的独立危险因素,早期检测miR-378a-3p有助于预测ACS的发病情况。     

Circle RNA hsa_circRNA_100290 serves as a ceRNA for miR-378a to regulate oral squamous cell carcinoma cells growth via Glucose transporter-1 (GLUT1) and glycolysis

Aerobic glycolysis (the Warburg effect) is a robust metabolic hallmark of most tumors, including oral squamous cell carcinoma (OSCC). Glucose transporter 1 (GLUT1), a major glucose transporter regulating the glucose uptake, is upregulated in OSCC and participated in the cell glycolysis of OSCC. The deregulation and function of noncoding RNAs in cancers have been widely reported. Reportedly, hsa_circular RNA (circRNA)_100290 (circ_SLC30A7) is significantly upregulated (fold change = 6.91, p < 0.0000001) in OSCC. According to online tools prediction (miRWalk, miRanda, and Targetscan), miR-378a could simultaneously target circRNA_100290 and GLUT1. Herein, the expression of circRNA_100290 and GLUT1 remarkably increased in oral tumor tissue specimens and cells. In OSCC cell lines, cell proliferation and glycolysis could be remarkably downregulated by circRNA_100290 silence, which could be rescued by GLUT1 overexpression. Conversely, miR-378a expression could be remarkably inhibited in tumor tissue specimens and cells. The effect of miR-378a overexpression on OSCC cells was similar to those of circRNA_100290 silence. miR-378a directly bound to circRNA_100290 and GLUT1 3′-untranslated region, circRNA_100290 could remarkably relieve miR-378a-induced inhibition on GLUT1 via acting as a competing endogenous RNA (ceRNA). miR-378a inhibition remarkably attenuated the effect of circRNA_100290 silence on cell proliferation and glycolysis in OSCC cell lines. In summary, circRNA_100290 serves as a ceRNA to counteract miR-378a-mediated GLUT1 suppression, thus promoting glycolysis and cell proliferation in OSCC. We provide a reliable experimental basis for understanding the mechanism of cell growth and glycolysis deregulation in OSCC.     

Involvement of circRNA_0007059 in the regulation of postmenopausal osteoporosis by promoting the microRNA-378/BMP-2 axis

Increasing evidence suggests that postmenopausal osteoporosis (PMO), a severe disturbance, imposes heavy physical, psychosocial, and financial burdens and dramatically influences the quality of life of postmenopausal women. Circular RNAs (circRNAs) and microRNAs (miRs) play important roles in the occurrence and development of PMO. However, the roles of circRNAs and miRs in osteoporosis regulation still need to be further investigated. circRNAs with different expression levels in patients with PMO were screened via RNA-seq and bioinformatics analysis. We found that circ_0007059 was upregulated in patients with PMO and during osteoclastogenesis of human bone marrow stromal cells (hBMSCs). Next, we investigated the effect of circ_0007059 overexpression during osteoclastogenesis of hBMSCs. circ_0007059 overexpression attenuated hBMSC differentiation into osteoclasts in vitro. This was demonstrated by downregulated bone morphogenetic protein 2 (BMP-2) expression, upregulated osteoclast-specific gene expression, and TRAP staining. circ_0007059 was demonstrated to directly target miR-378, which in turn targeted BMP-2 via bioinformatics analysis and the dual-luciferase reporter assay. Transfection of the miR-378 mimic reversed the effect of circ_0007059 on the osteoclastogenesis of hBMSCs. These results suggest that circ_0007059 plays an important role in osteoclastogenesis via the miR-378/BMP-2 signaling pathway. Targeting the circ_0007059/miR-378/BMP-2 axis is possibly a novel idea in osteoporosis treatment.