骨肉瘤细胞外泌体激活肺组织成纤维细胞炎性信号通路

谢显彪; 温丽丽; 吕东明; 李洪波; 刘伟海; 邹雨桐; 姚浩; 沈靖南

您当前的位置：

首页 >

文章列表页 >

骨肉瘤细胞外泌体激活肺组织成纤维细胞炎性信号通路

基础研究 | 更新时间：2023-11-01

- 骨肉瘤细胞外泌体激活肺组织成纤维细胞炎性信号通路
- Osteosarcoma Cells Derived Exosome Activate Inflammatory Signaling Pathwaysin Lung Fibroblast Cells
- 中山大学学报（医学科学版） 2020年41卷第4期页码：509-514
- 作者机构：
  
  1.中山大学附属第一医院骨肿瘤科，广东广州 510058
  2.广东省骨科学重点实验室，广东广州 510058
  3.中山大学肿瘤防治中心麻醉科，广东广州 510060
- 作者简介：
  
  谢显彪，副主任医师，研究方向：骨肿瘤，E-mail：xiexbiao@mail.sysu.edu.cn
- 基金信息：
  
  国家自然科学基金(81772864);中山大学青年教师培育项目(19ykpy198)
- DOI：
  中图分类号： R738
- 纸质出版日期：2020-07-15，
  
  收稿日期：2020-03-26，
扫描看全文
谢显彪,温丽丽,吕东明等.骨肉瘤细胞外泌体激活肺组织成纤维细胞炎性信号通路[J].中山大学学报（医学科学版）,2020,41(04):509-514.

XIE Xian-biao,WEN Li-li,LV Dong-ming,et al.Osteosarcoma Cells Derived Exosome Activate Inflammatory Signaling Pathwaysin Lung Fibroblast Cells[J].Journal of Sun Yat-sen University(Medical Sciences),2020,41(04):509-514.
谢显彪,温丽丽,吕东明等.骨肉瘤细胞外泌体激活肺组织成纤维细胞炎性信号通路[J].中山大学学报（医学科学版）,2020,41(04):509-514. DOI：

XIE Xian-biao,WEN Li-li,LV Dong-ming,et al.Osteosarcoma Cells Derived Exosome Activate Inflammatory Signaling Pathwaysin Lung Fibroblast Cells[J].Journal of Sun Yat-sen University(Medical Sciences),2020,41(04):509-514. DOI：

摘要

目的

本研究旨在研究骨肉瘤细胞来源外泌体对靶器官微环境的调控作用及其分子机制。

方法

提取骨肉瘤细胞143B外泌体并鉴定，通过PKH26荧光染色后经尾静脉注入裸鼠体内，24 h后提取肝、脾、肺、肾及脑组织，荧光显微镜下计数不同视野红色荧光数量；通过免疫荧光检测不同类型细胞中外泌体摄取情况；143B外泌体与人肺成纤维细胞共培养，荧光显微镜检测摄取情况，进一步利用qPCR检测炎性因子IL-1β、IL-6及TNF-α表达水平，通过蛋白印迹法检测炎性信号通路NF-κB通路p-p65和MAPK通路p-ERK及p-p38的变化。

结果

143B细胞外泌体中TSG101、Flotillin-1、CD63、CD9表达，Calnexin表达缺失(

0.05)；电镜外泌体呈“茶托”样形状，大小为(141.92±52.85) nm；外泌体经裸鼠尾静脉注射后进入肺组织含量显著高于肝、肾、脾、脑等器官（

0.05）；外泌体与人肺成纤维细胞共培养后炎性因子IL-1β、IL-6及TNF-α mRNA水平明显升高（

0.05），同时p-p65，p-ERK及p-p38均显著上调（

0.05）。

结论

骨肉瘤细胞外泌体可激活肺成纤维细胞NF-κB及MAPK炎性信号通路，进而促进炎性因子IL-1β、IL-6及TNF-α产生。

Abstract

Objective

To investigate the function and molecular mechanism of osteosarcoma cells derived exosome on microenvironment of target organs.

Methods

The osteosarcoma derived exosomes were extracted and injected into nude mice through tail vein after PKH26 fluorescence staining. The liver

spleen

lung

kidney and brain tissues were extracted 24 hours later and then the amount of red fluorescence in different fields was counted under fluorescence microscope. The uptake of exosomes in different types of cells was detected by immunofluorescence. 143B derived exosomes were co-cultured with human lung fibroblasts

and the uptake was detected by fluorescence microscopy. The expression levels of inflammatory cytokines IL-1β

IL-6 and TNF-α were detected by RT-qPCR

while the changes of p-p65 in inflammatory signaling pathway of NF-κB and p-ERK

p-p38 in MAPK signaling were detected by western blotting.

Results

TSG101

Flotillin-1

CD63 and CD9 were expressed in 143B derived exosomes

and Calnexin expression was absent (

0.05). The exosomes presented a saucer-like structure under electron microscope. The size of the exosomes is (141.92±52.85) nm. The exosomes distributed more in lung tissue than liver

kidney

spleen and brain after injection through the tail vein of nude mice (

0.05). The mRNA levels of inflammatory cytokines IL-1β

IL-6and TNF-α were significantly increased in human lung fibroblast cells after incubation with 143B exosomes (

0.05). p-p65

p-ERK and p-p38MAPK were significantly up-regulated (

0.05).

Conclusions

Osteosarcoma cells derived exosomes could activate inflammatory signaling pathway NF-κB and MAPK

and up-regulate the expression of the inflammatory cytokines IL-1β

IL-6 and TNF-α in lung fibroblast cells.

关键词

外泌体骨肉瘤成纤维细胞炎性信号通路

Keywords

exosomeosteosarcomafibroblast cellinflammatory microenvironment

references

Chou AJ， Geller DS， Gorlick R. Therapy for osteosarcoma： where do we go from here？［J］. Paediatr Drugs， 2008， 10（5）： 315-327.

Briccoli A， Rocca M， Salone M， et al. High grade osteosarcoma of the extremities metastatic to the lung： long-term results in 323 patients treated combining surgery and chemotherapy， 1985-2005［J］. Surg Oncol， 2010， 19（4）： 193-199.

Wortzel I， Dror S， Kenific CM， et al. Exosome-mediated metastasis： communication from a distance［J］. Dev Cell， 2019， 49（3）： 347-360.

Liu Y， Cao X. Characteristics and significance of the pre-metastatic niche［J］. Cancer Cell， 2016， 30（5）： 668-681.

French KC， Antonyak MA， Cerione RA. Extracellular vesicle docking at the cellular port： Extracellular vesicle binding and uptake［J］. Semin Cell Dev Biol， 2017， 67： 48-55.

Becker A， Thakur BK， Weiss JM， et al. Extracellular vesicles in cancer： cell-to-cell mediators of metastasis［J］. Cancer Cell， 2016， 30（6）： 836-848.

Lobb RJ， Lima LG， Moller A. Exosomes： Key mediators of metastasis and pre-metastatic niche formation［J］. Semin Cell Dev Biol， 2017， 67： 3-10.

Rana S， Malinowska K， Zoller M. Exosomal tumor microRNA modulates premetastatic organ cells［J］. Neoplasia， 2013， 15（3）： 281-295.

黄向荣，张红河，来茂德. 外泌体来源的长链非编码RNA在肿瘤中的效应和机制［J］. 中华病理学杂志， 2019， 48（9）： 742-746.

Huang XR， Zhang HH， Lai MD. Effects and mechanisms of exosome⁃derived long non⁃coding RNAs in tumors［J］. Chin J Pathol， 2019， 48（9）： 742-746.

Hoshino A， Costa-Silva B， Shen TL， et al. Tumour exosome integrins determine organotropic metastasis［J］. Nature， 2015， 527（7578）： 329-335.

Fang T， Lv H， Lv G， et al. Tumor-derived exosomal miR-1247-3p induces cancer-associated fibroblast activation to foster lung metastasis of liver cancer［J］. Nat Commun， 2018， 9（1）： 191.

Tomita T， Sakurai Y， Ishibashi S， et al. Imbalance of Clara cell-mediated homeostatic inflammation is involved in lung metastasis［J］. Oncogene， 2011， 30（31）： 3429-3439.

Rutkowski MR， Stephen TL， Svoronos N， et al. Microbially driven TLR5-dependent signaling governs distal malignant progression through tumor-promoting inflammation［J］. Cancer Cell， 2015， 27（1）： 27-40.

Yan HH， Pickup M， Pang Y， et al. Gr-1+CD11b+ myeloid cells tip the balance of immune protection to tumor promotion in the premetastatic lung［J］. Cancer Res， 2010， 70（15）： 6139-6149.

赵宁，侯成成，章鹏，等. NOD样受体在类风湿关节炎患者外周血中的表达［J］.新医学， 2019， 50（11）： 855-858.

Zhao N， Hou CC， Zhang P， et al. Expression of NOD-like receptors in peripheral blood of patients with rheumatoid arthritis［J］. J New Med， 2019， 50（11）： 855-858.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

RNA结合蛋白PTBP1与磷酸化-AXL共表达在骨肉瘤中的临床意义

成纤维细胞与间充质干细胞来源外泌体修复急性皮肤创面的比较

基于高通量测序分析骨肉瘤发生和转移的分子特征

被α-synuclein激活的小胶质细胞通过外泌体转运miR-19a-3p抑制神经细胞自噬

聚乙二醇修饰脂质体阿霉素联合顺铂单次给药治疗晚期骨肉瘤的剂量递增试验