外泌体在卵巢癌诊断及治疗中的研究进展

摘 要：卵巢癌是常见的妇科肿瘤，其起病隐匿，临床诊断时往往已为中晚期，预后欠佳。外泌体是细胞分泌至胞外的一类含有生物学活性物质的小囊泡，它们在生理状态维持及疾病的进程中发挥重要作用。已有研究表明，外泌体在多种肿瘤的早期诊断以及治疗中起着重要作用，而在卵巢癌中的相关研究较少。本综述的目的是描述卵巢癌外泌体的基本特征及其在卵巢癌诊治中的新进展，从而为以后进一步研究卵巢癌的生物学特性提供参考。     

外泌体源miRNA在卵巢癌的发生和诊治中的研究进展

卵巢癌是女性常见的恶性肿瘤之一，其死亡率居妇科恶性肿瘤之首。患者就诊时通常已处于晚期，治疗效果差且易产生耐药性。故阐明卵巢癌发病的分子机制对于促进早期诊断和发现新的治疗方法至关重要。外泌体是一种携带多种物质的细胞外囊泡，包裹在外泌体中的miRNA广泛参与卵巢癌肿瘤微环境的形成、肿瘤的发生发展以及耐药性的产生，并在卵巢癌的诊治中具有较大的应用价值。本文就外泌体源miRNA在卵巢癌的发生和诊治中的研究进展进行综述。     

外泌体在肺癌诊断及治疗中的研究进展

肺癌是全球范围内发病率及死亡率最高的恶性肿瘤,其严重威胁着人类健康.外泌体(exosomes)是起源于多泡体的纳米级脂质膜囊泡,其内含有蛋白质、脂质、核酸等多种活性生物分子.外泌体在肺癌的发生与演进中发挥重要作用,其可促进肺癌微环境形成,增强肿瘤侵袭与转移能力,参与肿瘤免疫抑制及肿瘤放化疗抵抗,且对肺癌的早期诊断和治疗具有应用价值.本文将对外泌体在肺癌发生、发展、诊断及治疗中的研究进展进行综述.     

尿液外泌体在泌尿系统肿瘤中的研究进展

摘 要：外泌体是细胞主动分泌的具有脂质双分子层的小囊泡。尿液是在泌尿系统中产生,并存在大量的外泌体。肿瘤在发生、发展过程中能不断释放外泌体到细胞外。通过检测尿液中的外泌体可能早期追踪及监控泌尿系统肿瘤,代替活检穿刺或影像学等检测。外泌体还可用于脂溶性药物的装载、靶向药物及免疫疫苗的制备。全文对尿液外泌体的提取及其在泌尿系统肿瘤诊断、治疗的应用进展进行综述。     

循环血液外泌体miRNA在卵巢癌发生发展中作用及其诊断价值的研究进展

卵巢癌（ovarian cancer,OC）是女性恶性肿瘤死亡的主要原因。由于卵巢癌无症状发展,缺乏早期诊断标志物,大多数患者在晚期才被诊断出来。早期检测卵巢癌可显著提高总生存率,在过去的几十年里,微小RNA（miRNA）在癌症的发展中起着重要的作用,因此引起了极大的关注。miRNA可以在循环血液中稳定存在(如包裹在外泌体中),并可通过外泌体的分泌和转移在肿瘤细胞之间和肿瘤细胞微环境的沟通中发挥重要的作用。此外,外泌体miRNA在卵巢癌中的表达是失调的,可能反映肿瘤的恶性特征。因此评估外泌体来源的循环miRNA可能会为卵巢癌提供一类新的非侵袭性生物标志物。本综述概述了有关外泌体miRNA在卵巢癌发生发展过程中的作用以及循环血液外泌体miRNA作为卵巢癌早期诊断潜在生物标志物的现状。     

Exosome在肿瘤发展中的研究进展

外泌体是细胞分泌到细胞外的微小囊泡。近年来,越来越多的研究发现这些分泌到细胞外的微小囊泡会影响肿瘤进展。外泌体能够通过不同的机制参与肿瘤免疫、肿瘤侵袭与转移及化疗耐药等过程。外泌体标志物的研究将有可能作为肿瘤早期诊断、疗效评价或者预后判断的依据。本文就外泌体调控肿瘤进展的机制作一综述。     

外泌体microRNAs在卵巢癌诊断和治疗中作用的研究进展

外泌体microRNAs(miRNAs)在卵巢癌进展的各种过程中发挥重要作用，如卵巢癌细胞增殖、凋亡和侵袭。外泌体miRNAs通过各种途径参与基因表达、信使RNA合成、蛋白生成等，有助于对卵巢癌发病机制的理解。由于miRNAs在血浆外泌体中的有效性，miRNAs可作为新兴的癌症早期检测和治疗评估生物标志物。外泌体miRNAs作为一种信使，在肿瘤微环境中起着重要的作用，越来越多的研究也证实了miRNAs作为靶点治疗卵巢癌的可行性。     

外泌体在卵巢癌诊疗及转移中的研究进展

卵巢癌是妇科肿瘤领域内的一个热点主题。该种恶性肿瘤疾病的发病率高且隐匿,缺乏有效的早期发现及时诊断并治疗的方式,病情容易加重发生远处转移。因此罹患卵巢癌的女性5年生存率比较低。外泌体属于一类典型的囊性泡样的小体,参与肿瘤的发病及转移机制且在肿瘤疾病的治疗中起着重要作用。本篇综述主要介绍癌细胞外泌体在卵巢恶性肿瘤疾病的诊断、转移及治疗中的研究现状。     

外泌体在胃癌发生发展及转移中作用的研究现状及进展

外泌体是一种由各种细胞分泌的、广泛存在于人体各种体液中的直径为50~100 nm的细胞外囊泡,其囊泡内携带 DNA、microRNA(miRNA)、蛋白质及脂质等多种生物活性物质,其通过细胞间的物质交换及信号传递作用参与多种生理病理过程。目前研究显示外泌体在胃癌的早期诊断、发生发展、转移、治疗、耐药及预后等方面均发挥着重要作用。本文就外泌体在胃癌的发生发展及转移方面的研究现状及进展作一综述。     

外泌体对头颈鳞癌肿瘤微环境的调节作用

外泌体（exosome）是直径在30~150 nm具有脂质双层膜结构的细胞外囊泡,富含蛋白质、核酸、脂质等生物活性物质,通过介导细胞间通讯,在机体的生理和病理过程中发挥重要作用。已有研究表明外泌体与头颈鳞癌的发生发展密切相关,是头颈鳞癌早期诊断、靶向治疗和预后评价的潜在标志物。本文就外泌体及其在头颈鳞癌发生发展、血管形成、免疫逃逸、诊断治疗和预后评估中的研究进展进行综述,旨在探讨外泌体在头颈鳞癌的临床诊断和治疗中的潜在价值。     

Exosomes in development,metastasis and drug resistance of breast cancer

Transport through the cell membrane can be divided into active, passive and vesicular types (exosomes). Exosomes are nano‐sized vesicles released by a variety of cells. Emerging evidence shows that exosomes play a critical role in cancers. Exosomes mediate communication between stroma and cancer cells through the transfer of nucleic acid and proteins. It is demonstrated that the contents and the quantity of exosomes will change after occurrence of cancers. Over the last decade, growing attention has been paid to the role of exosomes in the development of breast cancer, the most life‐threatening cancer in women. Breast cancer could induce salivary glands to secret specific exosomes, which could be used as biomarkers in the diagnosis of early breast cancer. Exosome‐delivered nucleic acid and proteins partly facilitate the tumorigenesis, metastasis and resistance of breast cancer. Exosomes could also transmit anti‐cancer drugs outside breast cancer cells, therefore leading to drug resistance. However, exosomes are effective tools for transportation of anti‐cancer drugs with lower immunogenicity and toxicity. This is a promising way to establish a drug delivery system.     

Identification of prostate cancer biomarkers in urinary exosomes

Exosomes have recently appeared as a novel source of non-invasive cancer biomarkers since tumour-specific molecules can be found in exosomes isolated from biological fluids. We have here investigated the proteome of urinary exosomes by using mass spectrometry to identify proteins differentially expressed in prostate cancer patients compared to healthy male controls. In total, 15 control and 16 prostate cancer samples of urinary exosomes were analyzed. Importantly, 246 proteins were differentially expressed in the two groups. The majority of these proteins (221) were up-regulated in exosomes from prostate cancer patients. These proteins were analyzed according to specific criteria to create a focus list that contained 37 proteins. At 100% specificity, 17 of these proteins displayed individual sensitivities above 60%. Even though several of these proteins showed high sensitivity and specificity for prostate cancer as individual biomarkers, combining them in a multi-panel test has the potential for full differentiation of prostate cancer from non-disease controls. The highest sensitivity, 94%, was observed for transmembrane protein 256 (TM256; chromosome 17 open reading frame 61). LAMTOR proteins were also distinctly enriched with very high specificity for patient samples. TM256 and LAMTOR1 could be used to augment the sensitivity to 100%. Other prominent proteins were V-type proton ATPase 16 kDa proteolipid subunit (VATL), adipogenesis regulatory factor (ADIRF), and several Rab-class members and proteasomal proteins. In conclusion, this study clearly shows the potential of using urinary exosomes in the diagnosis and clinical management of prostate cancer.     

Induction of more aggressive tumoral phenotypes in LNCaP and PC3 cells by serum exosomes from prostate cancer patients

Prostate cancer (PCa) is the second most frequent and sixth most fatal cancer in men worldwide. Despite its high prevalence, our understanding of its etiology and the molecular mechanisms involved in the progression of the disease is substantially limited. In recent years, the potential participation of exosomes in this process has been suggested. Therefore, we aim to study the effect of exosomes isolated from the serum of patients with PCa on various cellular processes associated with increased tumor aggressiveness in two PCa cell lines: LNCaP-FGC and PC3. The exosomes were isolated by filtration wand ultracentrifugation. Their presence was confirmed by immunodetection of specific markers and their size distribution was analyzed by Dynamic Light Scattering (DLS). The results obtained demonstrated that serum exosomes from PCa patients increased migration of PC3 cells and neuroendocrine differentiation of LNCaP-FGC cells regardless of the grade of the tumor. PCa serum exosomes also enhanced the secretion of enzymes related to invasiveness and resistance to chemotherapeutics, such as extracellular matrix metalloproteases 2 and 9, and gamma-glutamyltransferase in both cell lines. Altogether, these findings support the pivotal participation of exosomes released by tumoral cells in the progression of PCa. Future studies on the molecular mechanisms involved in the observed changes could provide crucial information on this disease and help in the discovery of new therapeutic targets.     

iRGD‐modified exosomes effectively deliver CPT1A siRNA to colon cancer cells,reversing oxaliplatin resistance by regulating fatty acid oxidation

Fatty acid oxidation (FAO) plays a vital role in drug resistance in cancer cells. Carnitine palmitoyltransferase 1A (CPT1A), a key enzyme of FAO, is widely recognized as an emerging therapeutic target. Here, we confirmed that CPT1A was heterogeneously expressed in colon cancer cells, with a high expression in oxaliplatin‐resistant cells but low expression in oxaliplatin‐sensitive cells, and expression could be increased by oxaliplatin stimulation. In addition, we verified that CPT1A was more highly expressed in colon cancer tissues than in noncancerous tissues. Silencing CPT1A by siRNA or etomoxir, a specific small‐molecule inhibitor of CPT1A, could reverse the sensitivity of drug‐resistant colon cancer cells to oxaliplatin. Subsequently, the combination of oxaliplatin with CPT1A inhibition promoted apoptosis and inhibited proliferation. In addition, exosomes were generated with the iRGD peptide on the surface, which showed highly efficient targeting compared with control exosomes in vivo. Furthermore, we loaded and therapeutically applied iRGD‐modified exosomes with siCPT1A to specifically deliver siCPT1A into tumours to suppress FAO. As a consequence, iRGD‐modified exosomes showed the significant inhibition of CPT1A in tumour tissues and exhibited the ability to reverse oxaliplatin resistance and inhibit tumour growth by inhibiting FAO with high safety in vivo.     

Role of exosomes in the immune microenvironment of ovarian cancer

Exosomes are excretory vesicles that can deliver a variety of bioactive cargo molecules to the extracellular environment. Accumulating evidence demonstrates exosome participation in intercellular communication, immune response, inflammatory response and they even play an essential role in affecting the tumor immune microenvironment. The role of exosomes in the immune microenvironment of ovarian cancer is mainly divided into suppression and stimulation. On one hand exosomes can stimulate the innate and adaptive immune systems by activating dendritic cells (DCs), natural killer cells and T cells, allowing these immune cells exert an antitumorigenic effect. On the other hand, ovarian cancer-derived exosomes initiate cross-talk with immunosuppressive effector cells, which subsequently cause immune evasion; one of the hallmarks of cancer. Exosomes induce the polarization of macrophages in M2 phenotype and induce apoptosis of lymphocytes and DCs. Exosomes further activate additional immunosuppressive effector cells (myeloid-derived suppressor cells and regulatory T cells) that induce fibroblasts to differentiate into cancer-associated fibroblasts. Exosomes also induce the tumorigenicity of mesenchymal stem cells to exert additional immune suppression. Furthermore, besides mediating the intercellular communication, exosomes carry microRNAs (miRNAs), proteins and lipids to the tumor microenvironment, which collectively promotes ovarian cancer cells to proliferate, invade and tumors to metastasize. Studying proteins, lipids and miRNAs carried by exosomes could potentially be used as an early diagnostic marker of ovarian cancer for designing treatment strategies.     

卵巢癌腹水来源ｅｘｏｓｏｍｅｓ的免疫分子检测

目的　通过检测卵巢癌腹水中ｅｘｏｓｏｍｅｓ免疫相关分子的表达,探讨以ｅｘｏｓｏｍｅｓ为基础的肿瘤免疫治疗的可行性。方法　采用离心超滤联合蔗糖密度梯度离心法分离出４例恶性腹水中的ｅｘｏｓｏｍｅｓ,经透射电镜观察其超微结构。通过免疫胶体金技术和Ｗｅｓｔｅｒｎ杂交技术鉴定和确定ｅｘｏｓｏｍｅｓ的ＨＳＰ７０、ＨＬＡ-Ⅰ、ＨＬＡ-Ⅱ、ＮＹ-ＥＳＯ-１、ＭＡＧＥ-Ａ１及ＭＡＧＥ-Ｃ２等抗原分子表达。结果　卵巢癌患者恶性腹水中ｅｘｏｓｏｍｅｓ为直径３０～８０ｎｍ的膜性囊泡结构,圆形或椭圆形,腔内为低电子密度成分;经上述相应抗原的抗体 胶体金免疫电镜标记,囊外膜及腔内可见颗粒状电子致密物沉积。Ｗｅｓｔｅｒｎ杂交也证实上述抗原的存在,但在４例卵巢癌恶性腹水来源的ｅｘｏｓｏｍｅｓ中,各类免疫相关分子的表达谱或表达量是不同的。结论　卵巢癌腹水来源的ｅｘｏｓｏｍｅｓ富含各类肿瘤抗原、ＨＳＰ７０、ＨＬＡ-Ⅰ和ＨＬA-Ⅱ等免疫相关分子,并且在不同的卵巢个体存在明显的异质性,通过对腹水中ｅｘｏｓｏｍｅｓ免疫相关分子检测,有可能为以ｅｘｏｓｏｍｅｓ为基础的肿瘤免疫治疗提供帮助。     

Cancer exosomes trigger mesenchymal stem cell differentiation into pro-angiogenic and pro-invasive myofibroblasts

Stromal fibroblasts become altered in response to solid cancers, to exhibit myofibroblastic characteristics, with disease promoting influence. Infiltrating mesenchymal stem cells (MSC) may contribute towards these changes, but the factors secreted by cancer cells that impact MSC differentiation are poorly understood.We investigated the role of nano-metre sized vesicles (exosomes), secreted by prostate cancer cells, on the differentiation of bone-marrow MSC (BM-MSC), and the subsequent functional consequences of such changes. Purified exosomes impaired classical adipogenic differentiation, skewing differentiation towards alpha-smooth muscle actin (αSMA) positive myofibroblastic cells. A single exosomes treatment generated myofibroblasts secreting high levels of VEGF-A, HGF and matrix regulating factors (MMP-1, −3 and −13). Differentiated MSC had pro-angiogenic functions and enhanced tumour proliferation and invasivity assessed in a 3D co-culture model. Differentiation was dependent on exosomal-TGFβ, but soluble TGFβ at matched dose could not generate the same phenotype. Exosomes present in the cancer cell secretome were the principal factors driving this phenotype.Prostate cancer exosomes dominantly dictate a programme of MSC differentiation generating myofibroblasts with functional properties consistent with disease promotion.     

结直肠癌患者血清外泌体代替肿瘤组织检测K-Ras基因突变的研究*

目的 探讨结直肠癌患者血清外泌体代替肿瘤组织进行DNA中K-Ras基因12密码子突变检测的价值。方法 收集2014年5月至2015年9月于我院病理组织学诊断为局部晚期或转移性结直肠癌患者的外周血标本90例,使用ExoQuick试剂提取血清外泌体,盐析法提取外泌体及血细胞DNA,采用PCR方法扩增K-Ras基因12密码子并通过高通量测序法检测突变,比较血清外泌体DNA与血细胞DNA突变的特异性,比较外泌体与组织学检测的K-Ras基因12密码子突变情况。结果 90例结直肠癌患者中检测出外泌体K-Ras基因12密码子突变43例,突变率为47.8％。全组共检出4种突变类型,以G12D突变率为最高。与组织学检测比较,灵敏度为90.6％,一致性为81.1％（Kappa值=0.62,P＜0.05）。结论 血清外泌体DNA用于检测肿瘤相关突变,与肿瘤组织相比一致性较高,可作为液体活检的来源指导肿瘤的个体化治疗。     

Macrophage-secreted Exosomes Delivering miRNA-21 Inhibitor can Regulate BGC-823 Cell Proliferation

Exosomes, membranous nanovesicles, naturally carry bio-macromolecules or miRNA and play impoetantroles in tumor pathogenesis. Here, we showed that macrophages cell-derived exosomes can function as vehiclesto deliver exogenous miR-21 inhibitor into BGC-823 gastric cancer cells. Exosomes loaded with miR-21inhibitorsignificantly increased miR-21 levels in BGC-823, but miR-21inhibitor loaded in exosomes exerted an oppositeeffect. miRNA transfected with exosomes had less cellular toxicity to host cells compared to conventionaltransfection methods. The miR-21inhibitor loaded exosomes promoted the migration ability and reducedapoptosis of BGC-823 gastric cancer cells. These observations indicate that miR-21 acts as a tumor promoterby targeting the PDCD4 gene and preventing apoptosis of gastric cancer cells through inhibition of PDCD4expression. Furthermore, exosome -mediated miR-21 inhibitor delivery resulted in functionally more efficientinhibition and less cellular toxicity compared to conventional transfection methods. Similar approaches could beuseful in modification of target biomolecules in vitro and in vivo. These findings contribute to our understandingof the functions of miR-21 and exosomes as a carrier for therapy of gastric cancer.