The antitumor effect of TAT-DCF1 peptide in glioma cells
The antitumor effect of TAT-DCF1 peptide in glioma cells
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TAT-DCF1肽对胶质瘤细胞的抗肿瘤作用
DOI：
10.1016/j.npep.2018.06.004
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发表时间：
2018-10-01
期刊：
影响因子：
2.9
通讯作者：
Wen, Tieqiao
中科院分区：
文献类型：
--
作者：
Wang, Jiao;Wang, Qian;Wen, Tieqiao
背景胶质母细胞瘤是恶性程度最高的脑肿瘤之一，因此，建立有效的治疗方法至关重要。我们以前的研究结果表明，树突状细胞衍生因子（DCF 1）是一个有吸引力的候选人治疗胶质母细胞瘤，因为它的过度表达胶质母细胞瘤U251细胞导致凋亡。然而，这种递送方式限制了其临床应用，本文通过在大肠杆菌中表达TAT-DCF 1融合蛋白，以克服其目前递送方式存在的问题。DCF 1不同结构域的编码序列（全长，细胞质，细胞外，19个氨基酸），连同N-末端转录反式激活因子（达特）序列，扩增并亚克隆到细菌表达载体pET 30 a（+）中以产生（His）（6）-标记的融合蛋白。结果：免疫荧光和流式细胞仪检测表明，融合蛋白能有效地转导U251细胞。细胞活力、增殖和迁移试验表明，完整的TAT-DCF 1融合蛋白显著降低了U251的增殖和迁移。结论：TAT-DCF 1融合蛋白能有效地转导入胶质母细胞瘤U251细胞，并诱导其抗肿瘤作用，为进一步研究TAT-DCF 1的靶向性和药物的毒副作用提供了实验依据。
Background Glioblastoma is one of the most malignant brain cancer, thus, establishing an effective therapy is paramount. Our previous results indicate that dendritic cell-derived factor (DCF1) is an attractive candidate for therapy against Glioblastoma, since its overexpression in Glioblastoma U251 cells leads to apoptosis. However, the delivery approach limits its clinical application, in this paper, we expressed TAT-DCF1 fusion protein in E.coli in order to surmount its current delivery problems.Methods: The coding sequences of the different domains of DCF1 (full length, cytoplasmic, extracellular, 19-amino acid), together with the N-terminal transactivator of transcription (TAT) sequence, were amplified and subcloned into the bacterial expression vector pET30a(+) in order to produce (His)(6)-tagged fusion proteins. Coomassie blue-stained SDS-PAGE and Western blotting identification showed that purity of the fusion proteins.Results: Immunofluorescence and flow cytometry show that U251 cells were efficiently transduced with the fusion proteins. Cell viability, proliferation, and migration assays suggest that the complete TAT-DCF1 fusion protein significantly decreased U251 proliferation and migration. Flow cytometry further reveals that TAT-DCF1 triggered cellular apoptosis.Conclusions: In conclusion, these findings suggest that the TAT-DCF1 fusion protein was efficiently transduced into Glioblastoma U251 cells and induced the antitumor effect and support further investigation into specific targeting and side effects of TAT-DCF1 during drug delivery.