Gene therapy has become an important approach for treating cancer, and electroporation represents a technology for introducing therapeutic genes into a cell. An example of cancer gene therapy relying on gene electrotransfer is the use of immunomodulatory cytokines, such as interleukin 2 (IL-2) and 12 (IL-12), which directly stimulate immune cells at the tumour site. The aim of our study was to determine the effects of gene electrotransfer with two plasmids encoding IL-2 and IL-12 in vitro and in vivo. Two different pulse protocols, known as EP1 (600 V/cm, 5 ms, 1 Hz, 8 pulses) and EP2 (1300 V/cm, 100 µs, 1 Hz, 8 pulses), were assessed in vitro for application in subsequent in vivo experiments. In the in vivo experiment, gene electrotransfer of pIL-2 and pIL-12 using the EP1 protocol was performed in B16.F10 murine melanoma. Combined treatment of tumours using pIL2 and pIL12 induced significant tumour growth delay and 71% complete tumour regression. Furthermore, in tumours coexpressing IL-2 and IL-12, increased accumulation of dendritic cells and M1 macrophages was obtained along with the activation of proinflammatory signals, resulting in CD4 + and CD8 + T-lymphocyte recruitment and immune memory development in the mice. In conclusion, we demonstrated high antitumour efficacy of combined IL-2 and IL-12 gene electrotransfer protocols in low-immunogenicity murine B16.F10 melanoma.

基因治疗已成为治疗癌症的重要方法，而电穿孔代表了一种将治疗基因导入细胞的技术。依赖基因电转移的癌症基因治疗的一个例子是使用免疫调节细胞因子，如白介素 2 (IL-2) 和 12 (IL-12)，它们直接刺激肿瘤部位的免疫细胞。我们研究的目的是在体外和体内确定用两种编码 IL-2 和 IL-12 的质粒进行基因电转移的效果。体外评估了两种不同的脉冲协议，称为 EP1（600 V/cm、5 ms、1 Hz、8 个脉冲）和 EP2（1300 V/cm、100 µs、1 Hz、8 个脉冲），以用于随后的体内实验。在体内在实验中，使用 EP1 方案对 pIL-2 和 pIL-12 进行基因电转移在 B16.F10 鼠黑色素瘤中进行。使用 pIL2 和 pIL12 联合治疗肿瘤诱导显着的肿瘤生长延迟和 71% 的肿瘤完全消退。此外，在共表达 IL-2 和 IL-12 的肿瘤中，随着促炎信号的激活，树突状细胞和 M1 巨噬细胞的积累增加，导致小鼠 CD4 + 和 CD8 + T 淋巴细胞募集和免疫记忆发育。总之，我们证明了联合 IL-2 和 IL-12 基因电转移方案在低免疫原性小鼠 B16.F10 黑色素瘤中的高抗肿瘤功效。