Plasmid injection and application of electric pulses alter endogenous mRNA and protein expression in B16.F10 mouse melanomas.
Plasmid injection and application of electric pulses alter endogenous mRNA and protein expression in B16.F10 mouse melanomas.
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DOI：
10.1038/cgt.2010.43
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发表时间：
2010-12
影响因子：
6.4
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中科院分区：
文献类型：
--
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对组织施加电脉冲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中比较了8种有代表性的已发表方案传递质粒后报告基因的表达。F10小鼠黑色素瘤。该表达式根据用于输送的脉冲参数而显著变化。为了观察质粒注射和/或脉冲应用对内源性基因表达的可能影响，采用PCR阵列检测分娩后4和24小时应激相关mrna的水平。在质粒注射、单独电脉冲或联合使用时，观察到几种炎症趋化因子和细胞因子的mRNA水平升高。这种上调被单个实时反转录TaqMan PCR检测证实。在同一时间点从相同治疗的肿瘤中提取蛋白质，并通过ELISA和定制的多重头阵列检测炎症蛋白水平。炎症蛋白水平的升高通常与mRNA水平平行。观察到一些差异，这可能是由于不同的表达动力学。观察到的这些细胞因子和趋化因子的上调表达可能有助于或抑制基于免疫的癌症基因治疗的治疗效果。
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 four and 24 hours 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 ELISA 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.
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影响因子：
10.8
作者：
Dujardin, N;Staes, E;Préat, V
通讯作者：
Préat, V
影响因子：
12.4
作者：
Hartikka, J;Sukhu, I;Sawdey, M
通讯作者：
Sawdey, M
影响因子：
4.4
作者：
Ahlen, Gustaf;Soderholm, Jonas;Sallberg, Matti
通讯作者：
Sallberg, Matti
影响因子：
56.9
作者：
Fearon, DT;Locksley, RM
通讯作者：
Locksley, RM
影响因子：
5.1
作者：
Heller, L;Coppola, D
通讯作者：
Coppola, D