First explanations for differences in electrotransfection efficiency in vitro and in vivo using spheroid model
First explanations for differences in electrotransfection efficiency in vitro and in vivo using spheroid model
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DOI：
10.1016/j.ijpharm.2011.04.054
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发表时间：
2012-02-14
影响因子：
5.8
通讯作者：
Rols, M. -P.
中科院分区：
文献类型：
--
作者：
Chopinet, L.;Wasungu, L.;Rols, M. -P.
电基因疗法是一种很有前途的癌症治疗技术。然而，对电场作用下基因在肿瘤中的转移机制了解有限。而体外电转染是有效的，基因表达在肿瘤细胞在体内是弱的。为了确定这种差异的原因并揭示基因转移机制，我们建议使用多细胞肿瘤球体作为离体三维模型。比较悬浮细胞和球形细胞之间的效率，可以突出基本差异。在经典的电条件下（包括10个500 V/cm， 5 ms， 1 Hz的脉冲），悬浮细胞的转染率为23.75% +/- 2.450 SEM。在相同条件下，虽然编码GFP的质粒DNA与一半的电渗透细胞相互作用，但只有不到1%的细胞表达转基因。首先给出了体内电转染失败的答案：电场导致的细胞死亡是导致转染率低的原因，因为三维和多细胞结构阻止了DNA的传代。这些结果表明，球体是在体内繁殖的。验证球体作为电转染研究的相关模型，在体内实验之前打开了离体优化的可能性。(C) 2011 Elsevier B.V.版权所有
Electro-gene-therapy is a promising technique for cancer treatment. However, knowledge about mechanism of gene transfer with electric field in tumor is limited. Whereas in vitro electrotransfection is efficient, gene expression in tumoral cells in vivo is weak. To determine reasons for this difference and unravel gene transfer mechanisms, we propose to use multicellular tumor spheroid as a tridimensional model ex vivo. Comparison of efficiency between cell in suspension and cells in spheroid allow highlighting fundamental differences. For classical electrical conditions (consisting in 10 pulses of 500 V/cm, 5 ms, 1 Hz), suspension cells present a transfection rate of 23.75% +/- 2.450 SEM. In the same conditions on spheroid, although plasmid DNA coding GFP interact with half of electrically permeabilized cells, less than 1% of cells are expressing the transgene. First answers to in vivo electrotransfection failure are given: cell mortality due to electric field is responsible of this low transfection rate, as tridimensional and multicellular structure that prevents DNA passage. These results show that spheroid is reproducing in vivo situation. Validation of spheroid as a relevant model for electrotransfection study opens ex vivo optimization possibility before in vivo assay. (C) 2011 Elsevier B.V. All rights reserved.