Mahmoodzadeh Hosseini Hamideh, Soleimanirad Jafar, Mehdizadeh Aghdam Elnaz, Amin Mohsen, Imani Fooladi Abbas Ali
Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Med Oncol. 2015 Jan;32(1):409. doi: 10.1007/s12032-014-0409-6. Epub 2014 Dec 2.
Texosomes, nano-endosomal vesicles, are candidates for cancer immunotherapy due to their immunostimulating properties. We designed a new structure based on texosome and staphylococcal enterotoxin B (SEB) and assessed its cytotoxic impact on an ovarian cell line. Texosomes were isolated from tumor cells, and SEB was anchored onto by protein transfer method. MTT assay and Hoechst staining were used to identify the cytotoxic and apoptotic effects of this compound on treated cells with different concentrations of texosome-SEB (TEX-SEB). Moreover, the expression rate of bcl-2, bax, bak, bcl-xl and the activity of caspase-3 and caspase-9 were investigated. Treatments of the cells with 0.5, 2.5 and 10 μg/100 μl TEX-SEB were significantly cytotoxic within 24 h (p < 0.001). Hoechst staining revealed that all tested concentrations caused apoptosis after 24 h compared with the control cells (p < 0.001). Furthermore, it was found that treatment with all examined concentrations of TEX-SEB enhanced caspase-9 activity after 24 and 48 h, while caspase-3 activity was increased upon treatment with only 0.5 and 2.5 μg/100 μl of TEX-SEB after 24 h (p < 0.001). None of the concentrations of TEX-SEB affected the expression of the cancer-promoting genes. Our construct, the TEX-SEB, is a new model being able to create cytostatic properties on cancer cells.
外泌体,即纳米内体囊泡,因其免疫刺激特性而成为癌症免疫治疗的候选者。我们基于外泌体和葡萄球菌肠毒素B(SEB)设计了一种新结构,并评估了其对卵巢细胞系的细胞毒性影响。从肿瘤细胞中分离出外泌体,并通过蛋白质转移方法将SEB固定在外泌体上。采用MTT法和Hoechst染色来鉴定该化合物对不同浓度外泌体-SEB(TEX-SEB)处理的细胞的细胞毒性和凋亡作用。此外,还研究了bcl-2、bax、bak、bcl-xl的表达率以及caspase-3和caspase-9的活性。用0.5、2.5和10μg/100μl TEX-SEB处理细胞在24小时内具有显著的细胞毒性(p<0.001)。Hoechst染色显示,与对照细胞相比,所有测试浓度在24小时后均导致细胞凋亡(p<0.001)。此外,发现用所有检测浓度的TEX-SEB处理在24小时和48小时后增强了caspase-9的活性,而仅用0.5和2.5μg/100μl的TEX-SEB处理24小时后caspase-3的活性增加(p<0.001)。TEX-SEB的所有浓度均未影响促癌基因的表达。我们构建的TEX-SEB是一种能够对癌细胞产生细胞生长抑制特性的新模型。