Mahmoodzadeh Hosseini Hamideh, Ali Imani Fooladi Abbas, Soleimanirad Jafar, Reza Nourani Mohammad, Mahdavi Mehdi
Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
J BUON. 2014 Apr-Jun;19(2):440-8.
Exosomes (EXOs) are acellular vehicles used for cancer immunotherapy due to their immune-inducing properties. We synthesized a novel structure based on EXOs and staphylococcal enterotoxin B (SEB) and surveyed its cytostatic effect on a pancreatic cell line.
EXOs were purified from tumor cells and SEB was anchored on it by protein transfer method. To determine the cytotoxic and apoptosis-inducing effect of this structure, treated cells with different concentrations of EXO/SEB were examined by MTT assay and Hoechst staining method. In addition, the expression rate of bcl-2, bax, bak, fas, bcl-xl and the activity of caspase-3 and caspase-9 were assessed.
We observed that 0.5 and 2.5 μg/100μl of EXO/ SEB significantly (p<0.001) stimulated apoptosis after 24 hrs. The concentrations of 0.5 and 2.5μg/100μl of EXO/SEB raised the expression rate of bax, bak, fas (p<0.001) but had no impact on bcl-2 and bcl-xl after 48 hrs. Furthermore, it was shown that 0.5, 2.5 and 5 μg/100μl of EXO/SEB only increased the activity of caspase-3 after 48 hrs (p<0.001).
Our designed structure, the EXO/SEB, is a novel model being able to induce apoptosis.
外泌体（EXOs）因其免疫诱导特性而被用作癌症免疫治疗的无细胞载体。我们基于外泌体和葡萄球菌肠毒素B（SEB）合成了一种新型结构，并研究了其对胰腺细胞系的细胞抑制作用。
从肿瘤细胞中纯化外泌体，并通过蛋白质转移方法将SEB锚定在其上。为了确定这种结构的细胞毒性和诱导凋亡作用，通过MTT法和Hoechst染色法检测用不同浓度的EXO/SEB处理的细胞。此外，评估bcl-2、bax、bak、fas、bcl-xl的表达率以及caspase-3和caspase-9的活性。
我们观察到，0.5和2.5μg/100μl的EXO/SEB在24小时后显著（p<0.001）刺激细胞凋亡。0.5和2.5μg/100μl的EXO/SEB浓度在48小时后提高了bax、bak、fas的表达率（p<0.001），但对bcl-2和bcl-xl没有影响。此外，结果显示，0.5、2.5和5μg/100μl的EXO/SEB仅在48小时后增加了caspase-3的活性（p<0.001）。
我们设计的结构EXO/SEB是一种能够诱导细胞凋亡的新型模型。
