Department of Medical Laboratory and Radiation Sciences, Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA 23508, USA.
Cancer Gene Ther. 2010 Dec;17(12):864-71. doi: 10.1038/cgt.2010.43. Epub 2010 Aug 13.
The application of electric pulses to tissues causes cell membrane destabilization, allowing exogenous molecules to enter the cells. This delivery technique can be used for plasmid gene therapy. Reporter gene expression after plasmid delivery with eight representative published protocols was compared in B16.F10 mouse melanoma tumors. This expression varied significantly based on the pulse parameters utilized for delivery. To observe the possible influence of plasmid injection and/or pulse application on endogenous gene expression, levels of stress-related mRNAs 4 and 24 h after delivery were determined by PCR array. Increases in mRNA levels for several inflammatory chemokines and cytokines were observed in response to plasmid injection, electric pulses alone or the combination. This upregulation was confirmed by individual real-time reverse transcription TaqMan PCR assays. Proteins were extracted at the same time points from identically treated tumors and inflammatory protein levels were assayed by enzyme-linked immunosorbent assay and by a custom multiplex bead array. Increases in inflammatory protein levels generally paralleled mRNA levels. Some differences were observed, which may have been due to differing expression kinetics. The observed upregulated expression of these cytokines and chemokines may aid or inhibit the therapeutic effectiveness of immune-based cancer gene therapies.
电脉冲应用于组织会导致细胞膜不稳定，使外源分子进入细胞。这种传递技术可用于质粒基因治疗。在 B16.F10 小鼠黑色素瘤肿瘤中，比较了 8 种有代表性的已发表方案中质粒传递后的报告基因表达。根据传递过程中使用的脉冲参数，这种表达有很大差异。为了观察质粒注射和/或脉冲应用对内源基因表达的可能影响，在传递后 4 小时和 24 小时通过 PCR 阵列确定与应激相关的 mRNA 水平。与质粒注射、单独电脉冲或两者结合后，观察到几种炎症趋化因子和细胞因子的 mRNA 水平增加。通过单独的实时逆转录 TaqMan PCR 检测证实了这种上调。在相同处理的肿瘤中同时提取蛋白质，并通过酶联免疫吸附测定法和定制的多重珠阵列测定炎症蛋白水平。炎症蛋白水平的升高通常与 mRNA 水平平行。观察到的这些细胞因子和趋化因子的表达上调可能有助于或抑制基于免疫的癌症基因治疗的治疗效果。
