5-脱氧杂氮胞苷修饰的肝癌细胞来源胞外体对淋巴细胞增殖功能的影响  

The Effects of 5-Aza-deoxycytidine Modified Exosomes Derived from Hepatoma Cell on the Lymphocytes Proliferations.

作　　者：张彩虹[1,2] 肖文华[1] 耿排力[2] 

机构地区：[1]解放军总医院第一附属医院,北京100048 [2]青海大学医学院免疫学教研室,西宁810010

出　　处：《医学研究杂志》2011年第6期42-45,共4页Journal of Medical Research

基　　金：首都医学发展科研基金资助项目(2005-3086)

摘　　要：目的研究经5-脱氧杂氮胞苷(5-Aza-deoxycytidine,5-Aza-CdR)处理的肝癌细胞HepG2分泌的胞外体(exosomes)对免疫细胞增殖能力的影响。方法采用离心超滤等方法将经5-Aza-CdR处理和未处理过的肝癌细胞系HepG2细胞分泌的exosomes分离和纯化。Ficoll淋巴细胞分层液分离外周血单个核细胞(PMBC);将实验分为exosomes实验组(加药组)、exosomes对照组(未加药组)、加药exosomes提取后上清组、未加药exosomes提取后上清组、空白对照组。H3-TdR掺入法检测PBMC细胞增殖状况。结果 exosomes对照组及其上清液对淋巴细胞增殖功能有明显的抑制作用,与空白对照组比较有显著性差异(P<0.05);而exosomes及其上清液实验组对淋巴细胞增殖功能的影响明显减弱,与exosomes和上清液对照组比较存在显著的统计学差异(P<0.05),而与空白对照组无统计学上的差异(P>0.05)。结论经5-Aza-CdR修饰的exosomes及其上清液显著改善HepG2来源的exosomes对淋巴增殖功能的抑制作用。Objective To investigate the effects of 5 - Aza - deocyeytidine （5 - Aza - CdR） modified exosomes derived from hepa- toma cell HepG2on the lymphocytes proliferations. Methods Exosomes derived from HepG2 cells with or without treatment by 5 - aza - CdR were isolated and purified using ultrafiltration centrifugation and sucrose gradient ultracentrifugation. The Ficoll reagent was used to i- solate peripheral blood mononuclear cells. The experiments were divided five groups as follows：5 - Aza - CdR modified exosomes group and its supernatant group ; exosomes control group and its supernatant group ; and negative control group. H3 - TdR was used to detect lym- phocytes proliferations. We added different reagents into the 96 orifices containing PBMC according to the groups. The cells were incuba- ted for 24 hours and the multiplication of PBMC was detected by H3 - TdR. Results The proliferation of PBMC was significantly sup- pressed in the exosomes control group and its supernatant group in comparison with negative control group （ P 〈 O. 05 ）. However, there was no any suppression effect to be found in 5 - Aza - CdR modified exosomes group and its supernatant group in comparison with negative control group （ P 〉 0.05 ）. Conclusion 5 - Aza - CdR modified exosomes and its supernatant can significantly improve suppression effect of lymphocyte proliferation by exosomes from hepatoma cell.

关 键 词：EXOSOMES 5-脱氧杂氮胞苷 外周血单个核细胞肝癌细胞 

分 类 号：R392[医药卫生—免疫学]