肾癌细胞来源exosome的制备和免疫相关蛋白组成的初步研究

张尧; 张家摸; 何百成; 罗春丽; 吴小候

doi:10.13406/j.cnki.cyxb.2010.01.040

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肾癌细胞来源exosome的制备和免疫相关蛋白组成的初步研究

基础研究 | 更新时间：2024-01-17

- 肾癌细胞来源exosome的制备和免疫相关蛋白组成的初步研究
- Separation and identification of exosomes derived from renalcancer cells and primary study of protein composition
- 重庆医科大学学报 2010年35卷第1期页码：37-39
- 作者机构：
  
  1. 重庆医科大学附属第一医院泌尿外科
  2. 重庆医科大学药学院药理教研室
  3. 重庆医科大学检验系
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.01.040
  中图分类号： R737.11
- 纸质出版日期：2010，
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[1]张尧,张家摸,何百成,罗春丽,吴小候.肾癌细胞来源exosome的制备和免疫相关蛋白组成的初步研究[J].重庆医科大学学报,2010,35(01):37-39.

ZHANG Yao, et al. Separation and identification of exosomes derived from renalcancer cells and primary study of protein composition[J]. Journal of Chongqing Medical University, 2010,35(1):37-39.
[1]张尧,张家摸,何百成,罗春丽,吴小候.肾癌细胞来源exosome的制备和免疫相关蛋白组成的初步研究[J].重庆医科大学学报,2010,35(01):37-39. DOI： 10.13406/j.cnki.cyxb.2010.01.040.

ZHANG Yao, et al. Separation and identification of exosomes derived from renalcancer cells and primary study of protein composition[J]. Journal of Chongqing Medical University, 2010,35(1):37-39. DOI： 10.13406/j.cnki.cyxb.2010.01.040.

摘要

目的:分离纯化肾癌细胞分泌的exosome

检测exosome免疫相关蛋白的组成

探讨其潜在的临床应用价值。方法:体外培养人肾癌细胞株RC-2。用超滤和蔗糖重水密度梯度超速离心法分离纯化exosome

透射电镜观察其形态

Western blot检测其免疫相关蛋白的组成。结果:用超滤和蔗糖重水密度梯度超速离心法成功分离出exosome。透射电镜下exosome为类圆碟形

具有双层膜结构

直径30

00nm。exosome表达肾癌特异性抗原G250、热休克蛋白70（Heat shock protein70

HSP70）和细胞间粘附分子（Intercellular Adhesion Molecule-1

ICAM-1）。结论:超滤和蔗糖重水密度梯度超速离心法可以分离纯化肾癌细胞分泌的exosome

exosome表达免疫相关蛋白G250、HSP70、ICAM-1。其负载肾癌特异性抗原

具有免疫原性

可望成为治疗肾癌的新疫苗。

Abstract

Objective:To separate and purify exosomes derived from renal cancer cells and detect their immune associated protein in order to investigate their potential clinical value.Methods:Exosomes derived from RC-2 cells culture supematants were separated and purified by ultrafiltration and sucrose gradient centrifugation

which were identified by transmission electron microscope.The expressions of HSP70

ICAM-1 and G250 were detected by western blot.Results:Exosomes were successfully purified by ultrafiltration and sucrose gradient centrifugation

which were 30～100 nm in diameter with typical saucer-shape morphology

and expressed HSP70

ICAM-1 and G250 antigen.Conclusion:Ultrafiltration and sucrose gradient ultracentrifugation can be used to purify renal cancer cells-secreted exosomes.Exosomes

expressing HSP70

ICAM-1 and G250

might be the novel vaccine for renal cell cancer.

关键词

肾细胞癌exosome免疫相关蛋白

Keywords

renal cell carcinomaExosomeImmune associated protein

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