基于聚多巴胺纳米粒子的荧光共振能量转移法检测microRNA

基于聚多巴胺纳米粒子(PDA NPs)对Cy5标记单链DNA(Cy5-ssDNA)探针的荧光猝灭效应以及脱氧核糖核酸酶Ⅰ(DNaseⅠ)选择性切割DNA/RNA杂合结构中单链DNA的特性,建立了一种用于微小核糖核酸(miRNA)检测的新型恒温信号放大方法.在优化的实验条件下,体系的相对荧光强度(FR)与miR-21浓度的对数值成正比;对miR-21检测的线性范围为10 pmol/L~100 nmol/L,检出限达7 pmol/L.血清加标实验结果表明,该方法可用于生理环境下miR-21的检测.     

Identification of common microRNA-mRNA regulatory biomodules in human epithelial cancer

The complex regulatory network between microRNAs and gene expression remains an unclear domain of active research. We proposed to address in part this complex regulation with a novel approach for the genome-wide identification of biomodules derived from paired microRNA and mRNA profiles, which could reveal correlations associated with a complex network of dys-regulation in human cancer. Two published expression datasets for 68 samples with 11 distinct types of epithelial cancers and 21 samples of normal tissues were used, containing microRNA expression and gene expression profiles, respectively. As results, the microRNA expression used jointly with mRNA expression can provide better classifiers of epithelial cancers against normal epithelial tissue than either dataset alone (P=1×10^–10, F test). We identified a combination of 6 microRNA-mRNA biomodules that optimally classified epithelial cancers from normal epithelial tissue (total accuracy = 93.3%; 95% confidence intervals: 86%–97%), using penalized logistic regression (PLR) algorithm and three-fold cross-validation. Three of these biomodules are individually sufficient to cluster epithelial cancers from normal tissue using mutual information distance. The biomodules contain 10 distinct microRNAs and 98 distinct genes, including well known tumor markers such as miR-15a, miR-30e, IRAK1, TGFBR2, DUSP16, CDC25B and PDCD2. In addition, there is a significant enrichment (Fisher’s exact test P=3×10^–10) between putative microRNA-target gene pairs reported in 5 microRNA target databases and the inversely correlated microRNA-mRNA pairs in the biomodules. Further, microRNAs and genes in the biomodules were found in abstracts mentioning epithelial cancers (Fisher’s Exact test, unadjusted P<0.05). Taken together, these results strongly suggest that the discovered microRNA-mRNA biomodules correspond to regulatory mechanisms common to human epithelial cancer samples. In conclusion, we developed and evaluated a novel comprehensive method to systematically identify, on a genome scale, microRNA-mRNA expression biomodules common to distinct cancers of the same tissue. These biomodules also comprise novel microRNA and genes as well as an imputed regulatory network, which may accelerate the work of cancer biologists as large regulatory maps of cancers can be drawn efficiently for hypothesis generation.     

Current advances in electrochemical genosensors for detecting microRNA cancer markers

The electrochemical microRNA sensors are considered efficient, simple, and inexpensive analytical tools for the early diagnosis of cancer biomarkers. To enhance the sensitivity of the electrochemical genosensors toward detection of microRNAs, several amplification strategies based mainly on nanomaterials, enzymes, and oligonucleotides are investigated and discussed. This review highlights the main current achievements regarding the new promising and sensitive strategies for genosensors’ development, thus allowing for miroRNA analysis at the attomolar level.     

纳米孔道单分子检测结直肠癌MicroRNAs

MicroRNA（miRNA）可用于癌症的早期诊断、预后判断,其分析检测具有重要临床意义.结直肠癌的发生、发展与miRNA 21、miRNA 92等的异常表达明显相关.本研究设计了以poly（dT）_n为引导链的DNA探针（probe）并尝试使用α-溶血素（α-HL）纳米孔道单分子检测方法检测结直肠癌miRNAs.miRNA·probe复合物分子穿过α-HL纳米孔道限域空间时,由于probe链长、序列不同导致probe-α-HL相互作用不同,miRNA 92·probe 92、miRNA 21·probe 21、miRNA 16·probe 16输出为形态、阻断时间不同的多台阶特征信号,实现了三种miRNAs的有效区分.实验证明,此方法可以用于检测血清实际样品.因此,未来有望使用α-HL构建miRNA超灵敏单分子生物传感器.     

microRNA对肿瘤细胞增殖与分化的调控

microRNA(miRNA)是一种长度约为22核苷酸(nt)的非编码RNA,其主要通过碱基互补与靶mRNA的3'端非翻译区(3'UTR)结合,导致靶mRNA降解或抑制蛋白质的合成,在转录后水平调节基因的表达.miRNA突变、缺失或表达水平的异常会导致生理的异常与疾病的发生,与人类肿瘤疾病密切相关,它具有类似于癌基因或抑癌基因的作用,可参与肿瘤细胞的增殖、分化和细胞凋亡等调控过程.miRNA在肿瘤诊断和治疗方面具有广阔的应用前景.     

RNA药物抗乙型肝炎病毒的研究进展

乙型肝炎病毒(hepatitis Bvirus,HBV)导致的慢性感染已经成为威胁全球健康的主要问题之一.传统的抗乙肝药物如以拉米夫定和阿德福韦为代表的核苷类似物以及干扰素类药物在临床上都存在一定的局限性,因此寻找新的抗乙肝药物已经成为当务之急.本文主要从小干扰RNA(small interfering RNA,siRNA)、微小RNA(microRNA,miRNA)和配体指数富集的系统进化技术(systematic evolution of ligands by exponential enrichment,SELEX)筛选的RNA适配子这3个方面来分别阐述用小分子RNA药物抗HBV的研究进展和展望.     

Intrinsic Electrocatalysis of RNA as a Label‐free and Reagent‐less Tool for Detection of MicroRNAs

We show for the first time that RNA catalyzes hydrogen evolution reaction at mercury‐containing electrodes. We previously showed that DNA is electrocatalytically active, and so we compared heights and potentials of the RNA chronopotentimetric stripping (CPS) peaks with analogous signals of DNA of the same sequence and found out they were very similar. RNA peaks showed differences depending on the RNA base composition. Catalytic nature of CPS peak enabled detection of 25 pM microRNA in electrochemical cell, or 500 pM microRNA in a 5 μL solution drop (corresponding to 2.5 fmole of microRNA). This finding opens the door for simple, label‐free and reagent‐less analysis of low concentrations of RNA molecules.     

The epigenetic involvement in plant hormone signaling

The biosynthesis and signaling of plant hormones play a critical role in almost all biological processes. It is well-documented that phytohormones cross-talk with each other. Epigenetic mechanisms were suggested to regulate expression of downstream targets in hormone signaling pathways that help implement hormone functions. This new layer of complexities that integrate epigenetic information such as DNA methylation, chromatin remodeling, histone modification, microRNAs and siRNAs with plant hormone signaling and regulations of gene expression, has been gradually revealed. In this short review, the author tries to assemble recent progress to establish a molecular linkage between these two large and momentum research fields and also to help readers digest the literature.     

Modelling of glioblastoma growth by linking a molecular interaction network with an agent-based model

In this work, a mathematical model of malignant brain tumour growth is presented. In particular, the growth of glioblastoma is investigated on the intracellular and intercellular scale.

The Go or Grow principle of tumour cells states that tumour cells either migrate or proliferate. For glioblastoma, microRNA-451 has been shown to be an energy dependent key regulator of the LKB1 (liver kinase B1) and AMPK (AMP-activated protein kinase) pathway that influences the signalling for migration or cell division.

We introduce a mathematical model that reproduces these biological processes. The intracellular molecular interaction network is represented by a system of nine ordinary differential equations. This is put into a multiscale context by applying an agent-based approach: each cell is equipped with this interaction network and additional rules to determine its new phenotype as either migrating, proliferating or quiescent.

The evaluation of the proposed model by comparison of the results with in vitro experiments indicates its validity.