Separation and identification of the Exosomes derived from heat shocked mouse hepatoma cell line(H22) and preliminary investigation of its protein composition

YANG Lin; et al

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Separation and identification of the Exosomes derived from heat shocked mouse hepatoma cell line(H22) and preliminary investigation of its protein composition

更新时间：2024-01-17

- Separation and identification of the Exosomes derived from heat shocked mouse hepatoma cell line(H22) and preliminary investigation of its protein composition
- Journal of Chongqing Medical University Issue 6, Pages: 669-672(2008)
- 作者机构：
  
  1. 重庆医科大学基础医学院病理生理学教研室
  2. 重庆医科大学基础医学院病理生理学教研室,重庆,400016
- 作者简介：
- 基金信息：
- DOI：
  CLC： R735.7
- Published：2008，
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[1]杨麟,沈宜,李静,陈黎.热休克小鼠肝癌细胞(H_(22))源Exosomes的制备及其蛋白组成的初步研究[J].重庆医科大学学报,2008(06):669-672.

YANG Lin, et al. Separation and identification of the Exosomes derived from heat shocked mouse hepatoma cell line(H22) and preliminary investigation of its protein composition. [J]. Journal of Chongqing Medical University (6):669-672(2008)
[1]杨麟,沈宜,李静,陈黎.热休克小鼠肝癌细胞(H_(22))源Exosomes的制备及其蛋白组成的初步研究[J].重庆医科大学学报,2008(06):669-672. DOI：

YANG Lin, et al. Separation and identification of the Exosomes derived from heat shocked mouse hepatoma cell line(H22) and preliminary investigation of its protein composition. [J]. Journal of Chongqing Medical University (6):669-672(2008) DOI：

摘要

目的:探讨热休克对小鼠肝癌细胞H22分泌的Exosomes产量及蛋白组分的影响

为进一步研究Exosomes在生产和免疫治疗方面的改进提供理论依据。方法:经超速分级离心和蔗糖密度梯度离心纯化得到H22细胞分泌的Exosomes和经热休克处理后H22细胞分泌的Exosomes（Heat shocked Exosomes

HS-Exo）

透射电镜观察其形态

SDS-PAGE电泳分析其蛋白带组成

WesternBlot检测两者所含有的免疫分子情况。结果:在透射电镜下

HS-Exo与Exosomes形态相似

直径约20

0nm的膜性囊泡

结构完整;经热休克处理后小鼠肝癌细胞H22分泌的Exosomes产量增多

且在HS-Exo中表达的热休克蛋白70（HSP70）、热休克同源蛋白70（HSC70）、热休克蛋白60（HSP60）、粘附相关蛋白（ICAM-1）与在Exosomes中的表达相比显著上调（P

0.05）。结论:超速分级离心及蔗糖密度梯度离心提取纯化热休克小鼠肝癌细胞H22源的Exosomes的方法具有可行性;HS-Exo中HSP70、HSC70、HSP60、ICAM-1的表达比Exosomes更为丰富

提示HS-Exo可能比Exosomes具有更强的免疫活性。

Abstract

Objective:To study the effect of heat shock on the production and protein composition of Exosomes derived from a mouse hepatoma cell line(H22) in order to provide theoretical basis for the improvement in production and immunol therapy of exosomes. Methods:Normal Exosomes and heat shocked Exosomes(HS-Exo) were purified by serial ultracentrifugation and sucrose density gradiant centrifugation

then were observed and identified by electron microscopy.The protein composition of Exosomes and HS-Exo were analysed by SDS-PAGE electrophoresis. And their immunological factors were detected by Western blot. Results:Under the electron microscopic view

HS-Exo had similar morphologic pattern to Exosomes

and the structure was complete

as vesicles with a diameter ranging from 20~90nm;The production was increased in heat shocked Exosomes.And the expressions of immunological factors such as HSP70

HSC70

HSP60

ICAM-1 in HS-Exo were significantly up-regulated than those in Exosomes. Conclusion:Serial ultracentrifugation and sucrose density gradiant centrifugation can be used to purify HS-Exo. HSP70

HSC70

HSP60

ICAM-1 were expressed more in HS-Exo than in Exosomes

which may suggest that Exosomes derived from heat-shocked H22 cells would have more effective immunostimulatory functions.

关键词

Exosomes肝癌细胞热休克肿瘤疫苗

Keywords

Exosomeshepatoma cellheat shocktumor vaccine

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Related Institution

Department of Pathophsiology,Institute of Stem Cell and Tissue Engineering,College of Basic Medicine, Chongqing Medical University

Department of Endocrinology，Guangdong Provincial People’s Hospital · Guangdong Academy of Medical Sciences

Department of Pathophsiology,Institute of Stem Cell and Tissue Engineering,College of Basic Medicine, Chongqing Medical University

Department of Obstetrics and Gynecology,Zhujiang Hospital,Southern Medical University

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