A novel cell-penetrating peptide TAT-A1 delivers siRNA into tumor cells selectively
A novel cell-penetrating peptide TAT-A1 delivers siRNA into tumor cells selectively
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DOI：
10.1016/j.biochi.2012.09.020
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发表时间：
2013-02-01
期刊：
影响因子：
3.9
通讯作者：
Ren, Fa Zheng
中科院分区：
文献类型：
--
作者：
Fang, Bing;Jiang, Lu;Ren, Fa Zheng
siRNA在抗肿瘤治疗中具有广阔的应用前景。主要挑战是缺乏肿瘤特异性细胞内递送。在这项研究中，一个6个氨基酸的肽（A1）具有高亲和力的血管内皮生长因子受体1（VEGFR 1）与细胞穿透肽（CPP）的达特连接，形成肿瘤选择性CPP。为了评价TAT-A1的肿瘤靶向穿透性质，通过流式细胞术测量TAT-A1的摄取。采用激光共聚焦显微镜检测肿瘤细胞和正常细胞共培养的体外选择性。TAT-A1的内化效率显著高于达特（p < 0.05）。当TAT-A1加入到共培养的肿瘤细胞和正常细胞中时，由于识别肿瘤细胞上过表达的VEGFR 1，TAT-A1选择性地渗透到肿瘤细胞中。TAT-A1介导的siRNA转染效果与Lipofectamine 2000相似，证明TAT-A1是一种高效的siRNA转染载体。TAT-A1介导的siRNA在mRNA和蛋白质水平上均具有敲除效应。这些结果表明，肿瘤靶向TAT-A1可以作为一个很好的载体特异性输送抗癌药物。(C)2012年由Elsevier Masson SAS出版。
siRNA is promising in anti-tumor therapy. The main challenge is lack of tumor-specific intracellular delivery. In this study, a 6 amino acids peptide (A1) with high affinity for vascular endothelial growth factor receptor-1 (VEGFR1) was conjugated with a cell penetrating peptide (CPP) TAT to form a tumor-selective CPP. To evaluate the tumor-targeted penetrate property of TAT-A1, the uptake of TAT-A1 was measured by flow cytometry. The selectivity in vitro was tested in co-cultured tumor cells and normal cells by laser confocal microscope. The internalization efficiency of TAT-A1 was significantly higher than that of TAT (p < 0.05). TAT-A1 penetrated into tumor cells selectively when added to co-cultured tumor cells and normal cells due to the recognition of VEGFR1 which is over-expressed on tumor cells. Furthermore, siRNA was successfully transferred by TAT-A1 into tumor cells in a similar way of Lipofectamine 2000, which was proved to be an efficient vector. The knockout effect of siRNA transferred by TAT-A1 was obtained at both mRNA and protein level. These results indicated that the tumor-targeted TAT-A1 can act as an excellent vehicle for specific delivery of anti-cancer agents. (C) 2012 Published by Elsevier Masson SAS.