微小RNA在新生儿缺血缺氧性脑病中的表达

微小RNA(miRNA)是一类高度保守的内源性小分子RNA。miRNA主要通过选择性结合mRNA调控基因表达。目前研究结果表明，中枢神经系统存在大量miRNA，并参与神经细胞的正常生长、发育，以及组织损伤修复、肿瘤发生、神经退行性变等多种病理、生理过程。笔者拟就新生儿缺血缺氧性脑病(HIE) miRNA谱系的最新研究进展进行阐述，探讨其miRNA特异性表达，对新生儿HIE诊断和预后判断的意义，旨在为该病的相关诊治研究提供参考。     

MicroRNA检测方法的研究进展

MicroRNAs（miRNAs）是一类内源性的小分子单链非编码RNA,通过调节基因的表达在许多生命活动中起重要作用。在一些疾病（如癌症和自身免疫性疾病）发生时,miRNAs的表达谱可能发生改变,所以在药物的安全性评价中,其有望成为诊断或预后生物标志物。因此,准确测定miRNA的表达对于其应用十分重要。对传统的RNA印记（Northernblotting）、微阵列（microarray）和实时定量PCR（qRT-PCR）以及一些新的miRNAs检测方法（如基于纳米材料的miRNAs检测、核酸扩增等技术）进行概述,并阐述了这些方法的优缺点。     

HPV: Molecular pathways and targets

Infection of high-risk human papillomaviruses (HPVs) is a prerequisite for the development of cervical carcinoma. HPV infections are also implicated in the development of other types of carcinomas. Chronic or persistent infection of HPV is essential but HPV alone is inadequate, additional endogenous or exogenous cues are needed along with HPV to induce cervical carcinogenesis. The strategies that high-risk HPVs have developed in differentiating epithelial cells to reach a DNA-synthesis competent state leading to tumorigenic transformation are basically due to overexpression of the E6 and E7 oncoproteins and the activation of diverse cellular regulatory or signaling pathways that are targeted by them. Moreover, the Wnt/β-catenin/Notch and phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathways are deregulated in various cancers, and have also been implicated in HPV-induced cancers. These are basically related to the “cancer hallmarks,” and include sustaining proliferative signals, the evasion of growth suppression and immune destruction, replicative immortality, inflammation, invasion, metastasis and angiogenesis, as well as genome instability, resisting cell death, and deregulation of cellular energetics. These information could eventually aid in identifying or developing new diagnostic, prognostic biomarkers, and may contribute to design more effective targeted therapeutics and treatment strategies. Although surgery, chemotherapy and radiotherapy can cure more than 90% of women with early stage cervical cancer, the recurrent and metastatic disease remains a major cause of cancer mortality. Numerous efforts have been made to design new drugs and develop gene therapies to treat cervical cancer. In recent years, research on treatment strategies has proposed several options, including the role of HPV E5, E6, and E7 oncogenes, which are retained and overexpressed in most of the cervical cancers and whose respective oncoproteins are critical to the induction and maintenance of the malignant phenotype. Other efforts have been focused on antitumor immunotherapy strategies. It is known that during the development of cervical cancer, a cascade of abnormal events is induced, including disruption of cell cycle control, perturbation of antitumor immune response, alteration of gene expression, deregulation of microRNA and cancer stem cell and stemness related markers expression could serve as novel molecular targets for reliable diagnosis and treatment of HPV-positive cancers. However, the search for new proposals for disease control and prevention has brought new findings and approaches in the context of molecular biology indicating innovations and perspectives in the early detection and prevention of the disease. Thus, in this article, we discuss molecular signaling pathways activated by HPV and potential targets or biomarkers for early detection or prevention and the treatment of HPV-associated cancers.     

MicroRNA-326 sensitizes human glioblastoma cells to curcumin via the SHH/GLI1 signaling pathway

Glioblastoma multiforme is the most malignant and common brain tumor in adults and is characterized by poor survival and high resistance to chemotherapy and radiotherapy. Among the new chemotherapy drugs, curcumin, a popular dietary supplement, has proven to have a potent anticancer effect on a variety of cancer cell types; however, it remains difficult to achieve a satisfactory therapeutic effect with curcumin using the traditional single-drug treatment. In this study, we found that expression of miR-326, a tumor suppressor microRNA in various tumor types, resulted in a marked increase of curcumin-induced cytotoxicity and apoptosis and a decrease of proliferation and migration in glioma cells. Moreover, we found that combination treatment of miR-326 and curcumin caused significant inhibition of the SHH/GLI1 pathway in glioma cells compared with either treatment alone, independent of p53 status. Furthermore, in vivo, the curcumin-induced increase in miR-326 expression altered the anti-glioma mechanism of this combination treatment, which further reduced tumor volume and prolonged the survival period compared to either treatment alone. Taken together, our data strongly support an important role for miR-326 in enhancing the chemosensitivity of glioma cells to curcumin.     

微小核糖核酸miR-370-5p对前列腺癌细胞增殖影响的研究

目的 探讨微小核糖核酸miR-370-5p对前列腺癌细胞系PC3和DU145细胞增殖的影响.方法 将前列腺癌细胞系PC3和DU145分为2组:阴性对照组-转染随机序列(dsCon-trol);实验组-转染miR-370-5p或miR-370-5p+siP21.实时荧光定量聚合酶链反应(qPCR)检测各组细胞中p21 mRNA的表达及前列腺癌细胞系中miR-370-5p的基础表达情况;蛋白质印迹法检测p21蛋白的表达;集落形成实验检测各组单个细胞克隆增殖情况;细胞增殖实验检测转染后各组细胞的增殖能力.结果 与正常前列腺上皮细胞(RWPE-1)比较,前列腺癌细胞系PC3和DU145中miR-370-5p表达下降;与阴性对照组相比,实验组PC3和DU145细胞中p21 mRNA的相对表达量分别提高了(2.457±0.392)倍和(1.844±0.295)倍.实验组PC3和DU145细胞中p21蛋白的相对表达分别为(0.52±0.09)和(0.63±0.14),阴性对照组为(0.18±0.06)和(0.24±0.05),两组比较,差异有统计学意义(P<0.05).阴性对照组和实验组中PC3和DU145细胞的集落形成数目分别为(0.281±0.024)、(0.084±0.016)、(0.293±0.017)和(0.310±0.041)、(0.088±0.019)、(0.300±0.032),实验组在转染miR-370-5p后,细胞集落形成数目均较阴性对照组少;而在miR-370-5p+siP21共转染后,与转染miR-370-5p组相比较,PC3和DU145细胞集落形成数目均显著恢复,差异均有统计学意义(P<0.05).细胞增殖实验结果显示,实验组转染miR-370-5p后,PC3和DU145细胞在48、72、96 h的存活情况(用吸光度OD值表示)分别为(0.395±0.040)、(0.691±0.042)、(0.874±0.045)和(0.437±0.044)、(0.700±0.051)、(0.875±0.052),与阴性对照组相比,实验组细胞增殖能力明显下降(P<0.05);miR-370-5p+siP21共转染后48、72、96 h,PC3和DU145细胞的OD值分别为(0.675±0.041)、(1.072±0.124)、(1.323±0.136)和(0.633±0.106)、(1.072±0.167)、(1.337±0.102),与转染miR-370-5p比较,差异均有统计学意义(P<0.05).结论 miR-370-5p能够通过上调p21蛋白的表达抑制前列腺癌细胞的增殖.     

冠心病患者血浆微小核糖核酸表达与血小板活化水平及其相关性

目的 探讨冠心病患者血浆微小核糖核酸(miR)表达与血小板活化水平及两者的相关性.方法 回顾性分析急性冠脉综合征患者30例(ACS组)、稳定性心绞痛患者30例(SAP组)、健康体检者30例(CT组)的临床资料.ACS组及SAP组患者入院第1天即予阿司匹林肠溶片及氯吡格雷片双联抗血小板治疗,连续7 d.比较治疗前后冠心病患者血浆miR-34a、miR-146a、miR-96、血小板活化依赖性颗粒表面膜蛋白(CD62p)、血小板膜糖蛋白Ⅱb/Ⅲa复合物纤维蛋白原受体(PAC-1)水平及血小板聚集率;分析冠心病患者miR表达水平与血小板活化指标之间的相关性.结果 治疗前,ACS组与SAP组miR-34a、miR-146a、miR-96、PAC-1、CD62p水平及血小板聚集率均高于CT组(P<0.05),且ACS组高于SAP组(P<0.05);治疗后,ACS组与SAP组各指标水平均较治疗前有所下降(P<0.05),但ACS组与SAP组仍高于CT组,且ACS组高于SAP组(P<0.05).结论 不同类型冠心病患者血浆miR-34a、miR-146a、miR-96表达及血小板活化水平均存在差异;miR-34a、miR-146a及miR-96水平可能通过参与血小板的产生或激活的过程而影响冠心病的发生发展.     

先心病中microRNA对DNA甲基化的调控及其应用前景

正先天性心脏病是最常见的出生缺陷,在活产婴儿中的总体发病率为0.8%~[1],居非感染原因死亡率第1位。先心病发病机制至今仍未完全阐明,目前认为主要是由于遗产因素、环境因素单独或者共同作用导致的。近年来,表观遗传学在先心病中的作用越来越被人们所认识。表观遗传学是指在DNA序列不变的情况下,可决定基因表达与否并可稳定遗传下去的调控方式,包括DNA甲基化、非编码RNA、基因组印记、染色质组蛋白修饰等。DNA甲基     

微小RNA在卵巢早衰中的研究概况及应用前景

微小RNA（microRNA,简写为miRNA）是一类真核生物内源性小分子单链RNA（核糖核酸）,长约21~25个核苷酸的非编码RNA,通过结合到mRNA的3’非编码区（3’UTR）,降解mRNA或者抑制其翻译过程,从而抑制转录后基因表达。研究发现miRNA在妇科肿瘤、多囊卵巢综合征、子宫内膜异位症等疾病均有表达且起到了重要作用,最新研究发现miRNA在卵巢早衰发生发展过程中起到了重要作用,本文对miRNA在卵巢早衰研究中的进展及应用前景做如下报道。