Kranjc Brezar Simona, Kranjc Matej, Čemažar Maja, Buček Simon, Serša Gregor, Miklavčič Damijan
Department of Experimental Oncology, Institute of Oncology Ljubljana, Zaloška 2, 1000 Ljubljana, Slovenia.
University of Ljubljana, Faculty of Electrical Engineering, Tržaška 25, 1000 Ljubljana, Slovenia.
Vaccines (Basel). 2020 Jan 27;8(1):49. doi: 10.3390/vaccines8010049.
The contactless high intensity pulsed electromagnetic field (HI-PEMF)-induced increase of cell membrane permeability is similar to conventional electroporation, with the important difference of inducing an electric field non-invasively by exposing a treated tissue to a time-varying magnetic field. Due to the limited number of studies in the field of electroporation induced by HI-PEMF, we designed experiments to explore the feasibility of such a contactless delivery technique for the gene electrotransfer of nucleic acids in tissues in vivo. By using HI-PEMF for gene electrotransfer, we silenced enhanced green fluorescent protein (EGFP) with siRNA molecules against EGFP in B16F10-EGFP tumors. Six days after the transfer, the fluorescent tumor area decreased by up to 39% as determined by fluorescence imaging in vivo. In addition, the silencing of EGFP to the same extent was confirmed at the mRNA and protein level. The results obtained in the in vivo mouse model demonstrate the potential use of HI-PEMF-induced cell permeabilization for gene therapy and DNA vaccination. Further studies are thus warranted to improve the equipment, optimize the protocols for gene transfer and the HI-PEMF parameters, and demonstrate the effects of HI-PEMF on a broader range of different normal and tumor tissues.
非接触式高强度脉冲电磁场(HI-PEMF)诱导的细胞膜通透性增加与传统电穿孔相似,重要的区别在于通过将处理过的组织暴露于随时间变化的磁场来无创地感应电场。由于HI-PEMF诱导电穿孔领域的研究数量有限,我们设计了实验来探索这种非接触式递送技术用于体内组织中核酸基因电转移的可行性。通过使用HI-PEMF进行基因电转移,我们在B16F10-EGFP肿瘤中用针对EGFP的siRNA分子沉默了增强型绿色荧光蛋白(EGFP)。转移后六天,通过体内荧光成像确定,荧光肿瘤面积减少了高达39%。此外,在mRNA和蛋白质水平上证实了EGFP在相同程度上的沉默。在体内小鼠模型中获得的结果证明了HI-PEMF诱导的细胞通透性在基因治疗和DNA疫苗接种中的潜在用途。因此,有必要进一步开展研究以改进设备,优化基因转移方案和HI-PEMF参数,并证明HI-PEMF对更广泛的不同正常和肿瘤组织的影响。