外泌体在宫颈癌形成及其诊疗中的作用

宫颈癌作为女性第2大恶性肿瘤,仍然是全球范围内的公共卫生问题.外泌体是活细胞主动分泌的一种具有脂质双分子层结构的纳米级囊泡,能够携带蛋白质、脂质、DNA和RNA(包括mRNA、miRNA、lncRNA和circRNA)等多种具有生物学活性的物质.作为新型的细胞间通讯分子,外泌体不仅参与细胞间正常的信息传递和物质交换等生...     

外泌体传递环状RNA介导肿瘤化疗耐药

环状RNA(circular RNA,circRNA)作为非编码RNA家族的重要成员,是一类共价闭环结构的单链RNA,没有多聚腺苷酸尾和5'-与-3'末端,显示出高度稳定性、丰富性和物种保守性等特点.近年来研究发现,circRNA与肿瘤化疗耐药、恶性进展等在内的多种生物学进程密切相关,发挥着极其重要的作用.外泌体是由机...     

外泌体环状RNA在泌尿系统肿瘤中的研究进展

外泌体是一种包含了复杂RNA和蛋白质的膜性囊泡，其主要来源于细胞内溶酶体微粒内陷形成的多囊泡体，经多囊泡体外膜与细胞膜融合后释放到胞外基质中。外泌体在肿瘤微环境中介导细胞间通讯，其功能取决于来源的细胞类型。环状RNA是一类由前体mRNA反向剪接生成的非编码RNA，在外泌体中富集且稳定表达。外泌体环状RNA在疾病中发挥了重要的调控作用，其作为肿瘤标志物和治疗靶点的临床应用前景与价值现已成为研究热点。本文就外泌体环状RNA在泌尿系统肿瘤中的研究进展作一综述。     

外泌体在IgA肾病中的作用

胞外囊泡是指从细胞膜上脱落或者由细胞分泌的双层膜结构的囊泡状小体,主要由外泌体、微囊泡和凋亡小体组成.外泌体是一种能被几乎所有细胞分泌的胞外囊泡,在细胞间物质和信息传递中起重要作用.IgA肾病(IgA nephropathy,IgAN)是最常见的原发性肾小球肾炎.大量研究显示,外泌体中含有的多种脂质、核酸和蛋白质等丰富...     

外泌体作为肿瘤标志物的研究进展

外泌体是细胞内源性囊泡样生物纳米级膜结构,直径大小在40～100 nm之间,可由各种类型的细胞分泌释放。外泌体具有许多功能,如蛋白质、mRNA、miRNA和脂类的细胞间运输和传递,以及抗原递呈,还可能具有致癌的能力。肿瘤细胞所分泌释放的外泌体在肿瘤的发生、发展以及迁移等生理和病理过程中发挥重要的作用。目前从肿瘤外泌体中寻找特异性标志物已成为肿瘤研究者重点关注的方向,对肿瘤早期诊断、疗效评价和预后分析具有重要的意义。就近年来外泌体在肿瘤研究和诊断中的研究进展进行了综述。     

肿瘤外泌体lncRNA在肿瘤微环境中的作用

外泌体是细胞分泌的纳米级囊泡,富含多种生物活性物质,是细胞间通讯的重要媒介.长非编码RNA(long non-coding RNAs,lncRNAs)可从多个方面影响肿瘤的发生发展,并能特异地分选入外泌体中.肿瘤微环境(tumor microenvironment,TME)是由肿瘤细胞及非肿瘤细胞(例如内皮细胞、免疫细...     

外泌体环状RNA在肿瘤微环境中的研究进展

外泌体是细胞分泌的30～150 nm的细胞外囊泡,在肿瘤微环境(tumor microenvironment,TME)中介导细胞间通讯.环状RNA (circular RNA,circRNAs)是一类由前体mRNA (precursor mRNA,pre-mRNA)反向剪接生成的非编码RNA(non-coding RNA,ncRNA),在外泌体中富集且表达稳定.本文主要讨论外泌体起源和circRNAs在外泌体中的分选调控机制,阐述外泌体circRNAs在肿瘤微环境各个阶段中的作用与机制,包括血管生成、EMT、耐药等.最后,本文探讨外泌体circRNAs作为肿瘤标志物和治疗靶点的临床应用前景与价值.     

外泌体在小RNA基因治疗中的研究进展

小RNA,包括小干扰RNA以及微小RNA,已成为多种疾病的潜在治疗药物。目前,小RNA的运输载体主要是病毒或者合成试剂。然而这类载体往往毒性高且特异性低。外泌体是由内源细胞分泌出来的天然纳米材料,本身能够穿越生物膜并在细胞间传递小RNA。以外泌体为基础的小RNA递送作为一种新的转运方式,能够克服低效率,低特异性以及免疫反应等缺陷,有望成为新型载体。本文简要论述了以外泌体为载体的小RNA递送系统在临床治疗研究中的前沿进展。     

外泌体miR-221在胃癌中的作用

外泌体是一种存在范围广泛、功能作用多样的膜性小囊泡,与肿瘤的发生发展紧密相关。外泌体miR-221为内源性非编码的RNA分子,在胃癌相关研究中极具价值。本综述主要介绍了外泌体的形成特点、功能作用以及外泌体miR-221与胃癌的相关作用机制和诊疗进程之间的联系,指出外泌体miR-221水平的高表达与胃癌的发生发展关系密切,提示其可作为潜在的生物标志物,为胃癌的早期诊断和治疗提供新的研究思路,以帮助临床解决胃癌这一难题。     

外泌体在肝纤维化中的作用

外泌体(exosomes)是一种能被大多数细胞分泌的微小膜泡,是具有脂质双层膜结构的细胞外囊泡。现认为外泌体是细胞外囊泡(extracellular vesicles, EVs)的一种亚群。研究表明,外泌体是细胞间信息传递的一种载体。肝脏既可以分泌外泌体,同时也是其他组织细胞产生的外泌体的作用靶点,且肝内与肝外来源的外泌体与肝纤维化的形成、发生、发展均有密切联系。本文主要就外泌体在肝纤维化相关疾病中的作用及外泌体与肝纤维化指标之间的关系进行综述。     

外泌体组成特征及其作为细胞通讯和分子标记的生物学作用

外泌体是由细胞分泌的直径在30～100 nm之间的微小囊泡状结构,内含来源于细胞相关的蛋白质与核苷酸等生物分子。外泌体可由几乎所有类型的细胞分泌,并且在组织细胞生理和病理情况下皆可持续分泌,存在于多种体液当中。目前,外泌体作为细胞间通讯的新途径和作为疾病诊断的生物标记方面取得瞩目的研究进展。本文从外泌体的组成特征及其生物学作用进行了综述,重点介绍了外泌体作为细胞通讯的新途径和内含的蛋白质和核苷酸作为一种新型的生物标记物在疾病诊断和临床方面的应用潜力,还对外泌体在生命科学研究领域的潜在作用及其存在的问题进行了展望。     

肺癌患者血浆外泌体蛋白质组分析

目的:探究肺癌患者与肺部良性结节病人血浆外泌体中蛋白质组的差异。方法:收集肺癌与肺部良性结节病人的术前血浆,随机选取肺腺癌病人血浆15例、肺鳞癌病人血浆10例作为实验组,肺部良性结节病人血浆5例作为对照组,采用超速离心法分离得到外泌体,定量质谱鉴定分析肺癌病人与肺部良性结节病人血浆外泌体蛋白质表达的差异性,SDS-PAGE和蛋白质免疫印迹法检验提取到的蛋白质。结果:共检测到血浆外泌体蛋白质253个,并初步确定其中18个在肺癌病人血浆外泌体中表达上调,8个在肺癌病人血浆外泌体中表达下调,免疫印迹法验证了上调蛋白质中的补体蛋白C4a。结论:鉴定出26个外泌体差异表达蛋白质,为肺癌早期诊断提供了新的候选分子。     

GPC1 exosome and its regulatory miRNAs are specific markers for the detection and target therapy of colorectal cancer

Colorectal cancer (CRC) is the second leading cause of cancer‐related deaths worldwide. However, a biomarker for a sensitive and simple diagnostic test and highly effective target therapy of CRC is still clinically unavailable. This study is to investigate the evidence and significance of plasma GPC1 positive exosomes as a biomarker of CRC. Results showed that GPC1⁺ exosomes were successfully isolated from tissues and plasma. The percentage of GPC1⁺ exosomes and the GPC1 protein expression in exosomes from tumour tissues and plasma of CRC patients before surgical treatment was significantly elevated compared to that in the peritumoural tissues and the plasma of healthy controls. miR‐96‐5p and miR‐149 expression in tumour tissues and plasma of CRC patients as well as in the GPC1⁺ exosomes from CRC patients were significantly decreased compared to that in the peritumoural tissues and the plasma of healthy controls. Two months after surgical treatment, levels of all tested markers significantly normalized. Overexpression of miR‐96‐5p and miR‐149 significantly decreased GPC1 expression in HT‐29 and HCT‐116 cells, xenograft tumours, plasma in mice bearing HT‐29 and HCT‐116 tumours, and the secretion of GPC1⁺ exosomes from the HT‐29 and HCT‐116 cells and xenograft tumours. Overexpression of miR‐96‐5p and miR‐149 significantly decreased cell viability and increased cell apoptosis in HT‐29 and HCT‐116 cells, and inhibited the growth of xenograft HT‐29 and HCT‐116 tumours. In conclusion, the increased plasma GPC1⁺ exosomes and reduced plasma miR‐96‐5p and miR‐149 expression are specific markers for the diagnosis of CRC and targets for the therapy of CRC.     

Cancer Liquid Biopsy Using Integrated Microfluidic Exosome Analysis Platforms

Liquid biopsies serve as both powerful noninvasive diagnostic tools for early cancer screening and prognostic tools for monitoring cancer progression and treatment efficacy. Exosomes are promising biomarkers for liquid biopsies, since these nano‐sized extracellular vesicles (EVs) enrich proteins, lipids, mRNAs, and miRNAs from cells of origin, including cancer cells. Although exosomes are abundantly present in various bodily fluids, conventional exosome isolation and detection methods that rely on benchtop equipment are time‐consuming, expensive, and involve complicated non‐portable procedures. As an alternative, recently developed microfluidic platforms can perform effective exosome separation and detection for liquid biopsies using a single device. Such methods offer advantages of integrity, speed, cost‐efficiency, and portability over conventional benchtop and early microfluidic‐based single‐functional methods which can only separate or detect exosomes separately. These advances have made exosome‐based point‐of‐care (POC) applications possible. This review outlines recent integrated microfluidic‐based exosomal detection strategies to guide future development of such devices for use in liquid biopsies for early cancer screening, prognostic monitoring, and other potential POC applications.     

真核生物核酸外切体(exosome)的研究进展

RNA的加工和降解是调控基因时空表达的重要步骤,在调节生物体的生长和发育过程中起着至关重要的作用.几乎所有的RNA都是从一条长的前体加工处理而来,形成成熟的RNA发挥功能,之后进行降解.RNA的降解需要5′-3′核酸外切酶、3′-5′核酸外切酶及核酸内切酶的参与.在真核细胞中,部分3′-5′核酸外切酶所进行的RNA降解依赖于一种称为核酸外切体(exosome)的复合物.该复合物由9个核心蛋白亚基组成,已有的证据表明,其广泛参与了动物、酵母及植物体中多种RNA的加工和降解过程.本文综述了真核生物中核酸外切体的研究进展,讨论了该复合体在RNA加工降解过程中的作用机制.     

Cancer biomarker discovery and translation: proteomics and beyond

Introduction: Cancer is often diagnosed at late stages when the chance of cure is relatively low and although research initiatives in oncology discover many potential cancer biomarkers, few transition to clinical applications. This review addresses the current landscape of cancer biomarker discovery and translation with a focus on proteomics and beyond.

Areas covered: The review examines proteomic and genomic techniques for cancer biomarker detection and outlines advantages and challenges of integrating multiple omics approaches to achieve optimal sensitivity and address tumor heterogeneity. This discussion is based on a systematic literature review and direct participation in translational studies.

Expert commentary: Identifying aggressive cancers early on requires improved sensitivity and implementation of biomarkers representative of tumor heterogeneity. During the last decade of genomic and proteomic research, significant advancements have been made in next generation sequencing and mass spectrometry techniques. This in turn has led to a dramatic increase in identification of potential genomic and proteomic cancer biomarkers. However, limited successes have been shown with translation of these discoveries into clinical practice. We believe that the integration of these omics approaches is the most promising molecular tool for comprehensive cancer evaluation, early detection and transition to Precision Medicine in oncology.     

Cellular and extracellular miRNAs are blood‐compartment‐specific diagnostic targets in sepsis

Septic shock is a common medical condition with a mortality approaching 50% where early diagnosis and treatment are of particular importance for patient survival. Novel biomarkers that serve as prompt indicators of sepsis are urgently needed. High‐throughput technologies assessing circulating microRNAs represent an important tool for biomarker identification, but the blood‐compartment specificity of these miRNAs has not yet been investigated. We characterized miRNA profiles from serum exosomes, total serum and blood cells (leukocytes, erythrocytes, platelets) of sepsis patients by next‐generation sequencing and RT‐qPCR (n = 3 × 22) and established differences in miRNA expression between blood compartments. In silico analysis was used to identify compartment‐specific signalling functions of differentially regulated miRNAs in sepsis‐relevant pathways. In septic shock, a total of 77 and 103 miRNAs were down‐ and up‐regulated, respectively. A majority of these regulated miRNAs (14 in serum, 32 in exosomes and 73 in blood cells) had not been previously associated with sepsis. We found a distinctly compartment‐specific regulation of miRNAs between sepsis patients and healthy volunteers. Blood cellular miR‐199b‐5p was identified as a potential early indicator for sepsis and septic shock. miR‐125b‐5p and miR‐26b‐5p were uniquely regulated in exosomes and serum, respectively, while one miRNA (miR‐27b‐3p) was present in all three compartments. The expression of sepsis‐associated miRNAs is compartment‐specific. Exosome‐derived miRNAs contribute significant information regarding sepsis diagnosis and survival prediction and could serve as newly identified targets for the development of novel sepsis biomarkers.     

Isolation and Quantification of MicroRNAs from Urinary Exosomes/Microvesicles for Biomarker Discovery

Recent studies indicate that microRNA (miRNA) is contained within exosome. Here we sought to optimize the methodologies for the isolation and quantification of urinary exosomal microRNA as a prelude to biomarker discovery studies. Exosomes were isolated through ultracentrifugation and characterized by immunoelectron microscopy. To determine the RNA was confined inside exosomes, the pellet was treated with RNase before RNA isolation. The minimum urine volume, storage conditions for exosomes and exosomal miRNA was evaluated. The presence of miRNAs in patients with various kidney diseases was validated with real-time PCR. The result shows that miRNAs extracted from the exosomal fraction were resistant to RNase digestion and with high quality confirmed by agarose electrophoresis. 16ml of urine was sufficient for miRNA isolation by absolute quantification with 4.15×10⁵ copies/ul for miR-200c. Exosomes was stable at 4℃ 24h for shipping before stored at -80℃ and was stable in urine when stored at -80°C for 12months. Exosomal miRNA was detectable despite 5 repeat freeze-thaw cycles. The detection of miRNA by quantitative PCR showed high reproducibility (>94% for intra-assay and >76% for inter-assay), high sensitivity (positive call 100% for CKD patients), broad dynamic range (8-log wide) and good linearity for quantification (R²>0.99). miR-29c and miR-200c showed different expression in different types of kidney disease. In summary, the presence of urinary exosomal miRNA was confirmed for patients with a diversity of chronic kidney disease. The conditions of urine collection, storage and miRNA detection determined in this study may be useful for future biomarker discovery efforts.     

循环miRNA 表达谱在肺癌早期诊断、判断预后和指导化疗中的应用

MicroRNA(miRNA)是一种高保守,长度大概21-23个核苷酸,非蛋白编码RNA,起着调节基因表达的作用。近年来有关miRNA与肺癌的关系已经得到证实,并且成为当前研究的热点。miRNA能整体调节基因表达,这使得miRNA表达谱在作为生物信号方面比蛋白编码基因更具有提示作用。最近发现miRNA以被保护的状态存在于循环血液中,这使得miRNA表达的发现具有非侵袭性、重现性以及易检测性。研究显示血浆miRNA表达谱可作为肺癌生物信号分子,在肺癌早期诊断、判断预后和指导化疗药物应用等方面具有重要作用。本文将对血浆miRNA与肺癌的研究进展,以及在肺癌早期诊断、判断预后和指导化疗药物应用等方面作一综述。