槟榔碱通过促进巨噬细胞分泌含miR-155-5p外泌体诱导人口腔成纤维细胞的活化

目的 探讨槟榔碱（ARE）通过调控巨噬细胞外泌体内miR-155-5p水平诱导人口腔黏膜成纤维细胞向成纤维表型转化的作用机制。方法 以人单核细胞系（THP-1）与人口腔黏膜成纤维细胞系（HOMF）为研究对象。实验设为空白对照组、阴性对照组、ARE刺激组、ARE刺激后THP-1培养上清刺激组（CS）、ARE刺激后THP-1外泌体刺激组（EXO）、ARE刺激后THP-1无外泌体上清刺激组（NES）、miR-155-5p过表达组（miR-155-5p mimics）、ARE刺激后EXO处理对照组（NC+EXO-ARE）和miR-155-5p抑制联合ARE刺激后EXO处理（miR-155-5p inhibitor EXO-ARE）组。以流式细胞术检测THP-1细胞极化情况。Transwell细胞迁移实验检测HOMF细胞迁移能力。DCFH-DA检测细胞氧化应激水平。qRT-PCR检测细胞α-SMA、I型胶原和SOCS1的mRNA转录水平。免疫印迹实验检测细胞α-SMA、I型胶原与SOCS1蛋白表达。结果 流式检测显示,相较于对照组,ARE组THP-1细胞由M0趋向M1极化。ARE对THP-1和H...     

细胞外囊泡微小RNA在动脉粥样硬化中的作用研究进展

细胞外囊泡(EV)是脂质双层包裹的微型囊泡,广泛存在于各种生物体液中,并可通过转运多种生物活性物质调节细胞的分子通路和生物学行为,特别是微小RNA(miRNA)。近年研究发现细胞外囊泡中的miRNA在动脉粥样硬化的发生和发展中发挥重要的调节作用。本文综述了动脉粥样硬化时细胞外囊泡miRNA转移的选择性及其在动脉粥样硬化发展过程中的调控作用和在诊断治疗中的潜在应用前景。     

Exosomes reveal the dual nature of radiotherapy in tumor immunology

Radioresistance is the potential cause of cancer metastasis and recurrence. Radiation‐induced changes in exosomes can partially explain the undesirable prognosis of radiotherapy (RT). Exosomes, newly discovered ways of cell communication, carry the characteristics of their origin, resulting in their diversity. Various exosomes in the tumor microenvironment exert different function in immune response. In this review, the dual effect of RT on the immune system was described, and the effect of radiotherapy on tumors via exosomes was explored. The molecules in exosomes after RT were described to play immunosuppressive and immunocompetent roles: immune‐related receptors and cell signaling molecules involved in both adaptive and innate immune system were present. CD69, TIGIT, TIM‐3, LAG‐3 and the tumor necrosis factor (TNF) family that signal to T cells were shown to be regulated by exosomes after irradiation. The change in innate immunity‐derived like receptors, Leukocyte Immunoglobin‐Like Receptors (LILR) was described, as well as B7‐H3, V‐domain containing Ig suppressor of T cell activation (VISTA), and CD155 on tumor cells. These changed molecules inhibit and activate the immune system through different mechanisms. By analyzing the relationship between exosome‐derived molecules and immunity, this review shows that radiotherapy can induce immunosuppression and immune clearance through exosomes, thereby treating tumors and improving patient prognosis.     

外泌体lncRNA在肿瘤中的研究进展

外泌体是肿瘤液态活检的明星分子,作为“胞间通讯”的信使可以在全身体液循环。随着科学家们对长链非编码RNA(lncRNA)的深入研究,发现lncRNA在表观遗传修饰、转录调控与转录后调控等多个层面发挥作用。在肿瘤发生、发展过程中,外泌体携带的lncRNA能够改变肿瘤微环境,介导肿瘤细胞增殖、转移和耐药,促进血管生成和介导缺氧信号等,在肿瘤诊断、疗效监测和预后评估等方面具有广阔的应用前景。该文总结外泌体lncRNA在肿瘤发生、发展中的作用,并对外泌体lncRNA在肿瘤诊断中的临床应用价值进行综述。     

牙源性干细胞来源外泌体的功能研究新进展

近年来关于间充质干细胞来源外泌体的研究已成为热点,但有关牙源性干细胞来源外泌体的研究相对较少。相比于其他间充质干细胞,牙源性干细胞因易获取,并具有更强的免疫调节功能和组织再生能力而成为学者们研究的新方向。文章总结了近几年牙源性干细胞来源外泌体功能研究新进展,主要就牙源性干细胞来源外泌体的炎症调节功能、促组织再生功能和神经保护功能3个方面做一阐述。     

食管癌细胞来源的外泌体对肿瘤细胞迁移及侵袭能力的影响及机制研究

目的 探讨食管癌KYSE410细胞来源的外泌体对肿瘤细胞迁移及侵袭能力的影响及其机制.方法 超速离心法分离食管癌KYSE410细胞培养上清中的外泌体;采用透射电镜及Western blotting观察所获得的外泌体的形态及其标志物;共聚焦显微镜观察KYSE410、KYSE510和YES2细胞摄取荧光染料标记的KYSE410外泌体;采用Transwell小室实验分析3种食管癌细胞的迁移及侵袭能力以及KYSE410外泌体对3种食管癌细胞迁移及侵袭能力的影响;Western blotting检测Wnt/β-catenin和PI3K/Akt信号通路相关蛋白的变化.结果 透射电镜下观察KYSE410外泌体具有膜结构,直径分布在30～100nm;3种食管癌细胞的迁移及侵袭能力从大到小依次为KYSE410、KYSE510、YES2;KYSE410外泌体能够促进KYSE410、KYSE510、YES2细胞迁移及侵袭能力并增强3种食管癌细胞中β-catenin和p-Akt的表达.结论 来源于高迁移及侵袭能力的食管癌KYSE410细胞的外泌体能够促进自身和低迁移及侵袭能力的食管癌KYSE510、YES2细胞的迁移和侵袭,并且可能通过激活Wnt/β-catenin和PI3K/Akt信号通路发挥上述作用.     

树突状细胞-肝癌细胞株HepG2融合细胞来源的exosome抗肝癌效应的实验研究

目的:以肝癌(HCC)患者树突状细胞(DC)与肝癌细胞HepG2之融合细胞为来源制备exosome,检测其体外诱导同源T细胞产生特异性抗HepG2的免疫效应。方法:以HCC患者外周血单个核细胞(PBMC)为来源,应用重组人粒细胞/巨噬细胞集落刺激因子(rhGM-CSF)、白细胞介素-4(rhlL-4)体外诱导培养DC;聚乙二醇融合DC与肝癌细胞HepG2,免疫磁珠分选DC-HepG2融合细胞,以纯化的融合细胞上清为来源,超滤及超高速密度梯度离心法制备exosome;透射电镜观察exosome形态,流式细胞术、ELISA等方法检测exo-some蛋白分子表达;以融合细胞来源的exosome直接刺激效应淋巴细胞,MTT法检测效应淋巴细胞对靶细胞HepG2的杀伤活性;ELISA法检测该效应淋巴细胞分泌IFN-γ水平。结果:DC-HepG2融合细胞可分泌ex-osome,该exosome同时含有DC的特征分子CD86(94.6%)和肝癌细胞HepG2的AFP抗原(40.7%),ELISA法亦证实exosome提取物中含有AFP蛋白;DC-HepG2来源的exosome活化的淋巴细胞(Exo-L)对靶细胞HepG2的杀伤率明显高于单纯淋巴细胞组(L)(P<0.05),Exo-L组对HepG2的杀伤率亦显著高于对其他两种靶细胞(肝癌细胞SMMC7721、白血病细胞K562)的杀伤率(P<0.05)。靶细胞为HepG2时,Exo-L组IFN-γ分泌量明显高于L组(P<0.05)。结论:以肝癌患者DC与肝癌细胞HepG2之融合细胞为来源可制备获得exosome瘤苗,该瘤苗无须DC存在,可直接激活自体淋巴细胞,产生HepG2抗原特异CTL,显示了比传统制备的exosome更强的功能,有望成为HCC免疫治疗的有效途径。     

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??Objective    To isolate and identify exosomes derived from stem cells from apical papilla. Methods??Stem cells from apical papilla ??SCAP?? were primarily cultured and identified. SCAP-derived exosomes were isolated by ultracentrifugation. The morphology was observed by transmission electron microscopy. The markers CD9 and Alix were detected by Western blotting. Results??SCAP could release exosomes??which showed a cup-like microvesicle structure??with a diameter of about 30-100 nm under electronic microscopy. The specific markers CD9 and Alix were positively expressed. Conclusion??The results show that exosomes can be successfully isolated from culture supernatants of SCAP by ultracentrifugation.