成纤维细胞系3T3细胞来源exosome对小鼠乳腺癌细胞增殖能力的影响

目的 观察小鼠成纤维细胞系3T3来源的外泌小体（exosome）对小鼠乳腺癌细胞4T1增殖能力的影响,并探索其中可能的机制。方法 PureExo Exosome提取试剂盒提取3T3细胞上清液中的exosome,按照不同浓度及时间作用于4T1细胞,CCK8法检测4T1细胞的增殖能力,BrdU/PI双掺入法测定细胞DNA合成及细胞周期;免疫印迹法（Western blot）及荧光定量实时PCR（qPCR）检测人表皮生长因子受体2（epidermal growth factor receptor-2, EGFR2,也称HER2）及下游PI3K/AKT信号转导通路相关蛋白的变化。利用HER2单克隆抗体靶向药物赫赛汀（Herceptin）,观察exosome是否影响4T1细胞对于Herceptin敏感度。结果 exosome处理组OD450吸光度值显著高于对照组（P<0.05）,细胞增殖及细胞周期进程加快。Western blot及qPCR实验提示随着exosome浓度的增加,HER2表达逐渐升高, AKT磷酸化水平增加。而同时给予exosome可明显增加4T1细胞对Herceptin的敏感度。结论 小鼠成纤维细胞系3T3来源exosome可促进小鼠乳腺癌细胞4T1增殖及周期进程,并且可能通过HER2激活其下游PI3K/AKT信号通路发挥上述作用。     

Tumor-derived exosomes: Nanovesicles made by cancer cells to promote cancer metastasis

Nanomedicine usually refers to nanoparticles that deliver the functional drugs and siRNAs to treat cancer. Recent research has suggested that cancer cells can also make nanoparticles that also deliver functional molecules in promoting cancer metastasis, which is the leading cause of various cancer mortalities. This nanoparticle is called tumor-derived vesicles, or better-known as tumor-derived exosomes (TEXs). TEXs are nanoscale membrane vesicles (30–140 nm) that are released continuously by various types of cancer cells and contain tumor-derived functional biomolecules, including lipids, proteins, and genetic molecules. These endogenous TEXs can interact with host immune cells and epithelial cells locally and systemically. More importantly, they can reprogram the recipient cells in favor of promoting metastasis through facilitating tumor cell local invasion, intravasation, immune evasion, extravasation, and survival and growth in distant organs. Growing evidence suggests that TEXs play a key role in cancer metastasis. Here, we will review the most recent findings of how cancer cells harness TEXs to promote cancer metastasis through modulating vascular permeability, suppressing systemic immune surveillance, and creating metastatic niches. We will also summarize recent research in targeting TEXs to treat cancer metastasis.     

星形胶质细胞外泌体对缺氧缺血神经元的保护作用

目的 探索星形胶质细胞外泌体对缺氧缺血神经元的影响。方法 体外培养大鼠星形胶质细胞,通过差速离心法获取细胞上清液中的外泌体。采用透射电镜、Nanosight和Western blot鉴定外泌体;采用BCA法检测外泌体蛋白浓度。体外培养大鼠神经元,分为对照组、外泌体组、氧糖剥夺（OGD）组和OGD+外泌体组（n=3）。OGD组和OGD+外泌体组给予无糖培养基和缺氧处理;外泌体组和OGD+外泌体组分别给予终浓度为22 μg/mL的外泌体处理,对照组和OGD组给予等体积PBS处理。采用ELISA法检测神经元乳酸脱氢酶（LDH）水平;TUNEL法检测神经元凋亡指数。结果 外泌体鉴定结果显示差速离心法提取的外泌体符合外泌体特点;与对照组和外泌体组相比,OGD组LDH值和凋亡指数显著增加（P < 0.05）;与OGD组相比,OGD+外泌体组LDH值和凋亡指数均显著降低（P < 0.05）。结论 星形胶质细胞来源的外泌体对神经元缺氧缺血损伤有保护作用。     

Emerging role of engineered exosomes in nonalcoholic fatty liver disease

Nonalcoholic fatty liver disease(NAFLD) is the most common chronic liver disease worldwide.NAFLD comprises a continuum of liver abnormalities from nonalcoholic fatty liver to nonalcoholic steatohepatitis,and can even lead to cirrhosis and liver cancer.However,a well-established treatment for NAFLD has yet to be identified.Exosomes have become an ideal drug delivery tool because of their high transmissibility,low immunogenicity,easy accessibility and targeting.Exosomes with specific modifications...     

Microglial Exosome miR-7239-3p Promotes Glioma Progression by Regulating Circadian Genes

Glioma-associated microglial cells, a key component of the tumor microenvironment, play an important role in glioma progression. In this study, the mouse glioma cell line GL261 and the mouse microglia cell line BV2 were chosen. First, circadian gene expression in glioma cells co-cultured with either M1 or M2 microglia was assessed and the exosomes of M2-polarized and unpolarized BV-2 microglia were extracted. Subsequently, we labeled the exosomes with PKH67 and treated GL261 cells with them to investigate the exosome distribution. GL261 cell phenotypes and related protein expression were used to explore the role of M2 microglial exosomes in gliomas. Then a specific miR-7239-3p inhibitor was added to verify miR-7239-3p functions. Finally, the mouse subcutaneous tumorigenic model was used to verify the tumorigenic effect of M2 microglial exosomes in vivo. Our results showed that in gliomas co-cultured with M2 microglia, the expression of the BMAL1 protein was decreased (P < 0.01), while the expression of the CLOCK protein was increased (P < 0.05); opposite results were obtained in gliomas co-cultured with M1 microglia. After treatment with M2 microglial exosomes, the apoptosis of GL261 cells decreased (P < 0.001), while the viability, proliferation, and migration of GL261 cells increased. Increased expression of N-cadherin and Vimentin, and decreased E-cadherin expression occurred upon treatment with M2 microglial exosomes. Addition of an miR-7239-3p inhibitor to M2 microglial exosomes reversed these results. In summary, we found that miR-7239-3p in the glioma microenvironment is recruited to glioma cells by exosomes and inhibits Bmal1 expression. M2 microglial exosomes promote the proliferation and migration of gliomas by regulating tumor-related protein expression and reducing apoptosis.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12264-020-00626-z.     

外泌体在甲状腺癌诊治中的研究进展

甲状腺癌(Thyroid Carcinoma,TC)是世界上发病率增长最快的癌症之一,寻找新型高特异性、高灵敏度、无创的早期诊断和术后监测标记物成为当下研究热点。外泌体是细胞间信息物质交流的重要载体,由于其高稳定性、低细胞毒性、低免疫原性和高膜通透性等特征,使其具有成为无创、新型的疾病诊断和治疗手段的潜能。近年来,外泌体在甲状腺癌非侵入性疾病诊断、监测、给药、治疗和预后评估等领域取得了实质性突破。论文综述了TC来源的外泌体在肿瘤中发生和发展、诊断监测以及治疗中的应用前景。     

间充质干细胞外泌体在骨关节炎治疗修复中的研究进展

骨关节炎(OA)是种以关节软骨退变为特征的疾病,其缺乏干预性药物,对于晚期OA患者往往通过关节置换术以维持生活质量。外泌体(exosomes)是一种由不同细胞分泌的细胞外囊泡,可以传递DNA、微小RNA(microRNA,miRNA)、mRNA、蛋白质等多种信息,并以此通过多种方式进行细胞间信号传递和功能调节。间充质干...     

小鼠胰岛微血管内皮细胞外泌体的提取和鉴定方法

目的：建立小鼠胰岛微血管内皮细胞来源的外泌体的提取和鉴定方法,为糖尿病胰岛微血管内皮受损的发病机制研究及重建胰岛微循环完整性的应用提供必要材料。方法选取小鼠胰岛微血管内皮细胞株（MS-1）细胞作为研究对象,取其培养上清液,经多步离心结合蔗糖／D2O垫超速离心方法获取提取物,然后采用透射电镜和Western blotting 法对提取物进行形态和蛋白成分分析,以确定其是否为外泌体。结果提取物呈形态均一的类圆形囊泡,有完整双层膜包绕,内部含有低电子密度物质。囊泡大小约为40～100 nm,可散在分布,也可聚集成团,提取物均阳性表达CD63、CD9和TSG101分子。结论经过形态和蛋白成分分析证实所提取到的物质正是MS-1细胞来源的外泌体,便于后续临床和基础研究工作的开展。     

MicroRNAs in the regulation of autophagy and their possible use in age-related macular degeneration therapy

Age-related macular degeneration (AMD) is a progressive sight-impairing disease of the elderly. The pathogenic mechanisms of AMD are not well understood although both genetic and many environmental factors have been associated with the development of AMD. One clinical hallmark of AMD is the detrimental aggregation of damaged proteins. Recently, it has been suggested that the weakening of autophagy clearance is an important mechanism in the pathogenesis of AMD. Autophagy is important in the removal of damaged or no longer needed cellular material and its recycling. A considerable number of autophagy-targeting microRNAs (miRNAs), small RNA molecules and epigenetic regulators have been found to be either up- or down-regulated in AMD patients and experimental models. The important role of autophagy-targeting miRNAs is supported by several studies and can open the prospect of the use of these miRNAs in the therapy for AMD.     

Exosome与肿瘤耐药的研究进展

Exosome是直径40～100 nm的微囊泡,内含蛋白质、mRNA、microRNA等生物活性物质.机体内几乎所有细胞都可分泌exosome.Exosome不仅是细胞-细胞间物质信息交流的通讯工具,还参与免疫调节、炎症反应、肿瘤发生和耐药等生理病理过程.该文就exosome与肿瘤耐药之间的关系作一综述.