Targeted gene delivery in tumor xenografts by the combination of ultrasound-targeted microbubble destruction and polyethylenimine to inhibit survivin gene expression and induce apoptosis.
Targeted gene delivery in tumor xenografts by the combination of ultrasound-targeted microbubble destruction and polyethylenimine to inhibit survivin gene expression and induce apoptosis.
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DOI：
10.1186/1756-9966-29-152
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发表时间：
2010-11-23
期刊：
影响因子：
--
通讯作者：
Qiu RX
中科院分区：
文献类型：
--
作者：
Chen ZY;Liang K;Qiu RX
无创性和组织特异性的基因转染技术在临床基因治疗中具有重要的应用价值。本研究旨在确定超声靶向微泡破坏（UTMD）与聚乙烯亚胺（PEI）联合应用是否能增强肿瘤异种移植物中基因的体内转染，并阐明通过这种新技术靶向人生存素的短发夹RNA（shRNA）干扰治疗的基因沉默和凋亡诱导作用。将两种不同的表达载体（pCMV-LUC和pSIREN）与PEI一起孵育以制备阳离子复合物（PEI/DNA），并通过凝胶阻滞测定来确认。将人宫颈癌（Hela）肿瘤种植于裸鼠两侧皮下。将PBS、质粒、质粒和SonoVue微泡、PEI/DNA和SonoVue微泡通过尾静脉给予荷瘤小鼠。一个肿瘤暴露于超声照射，而另一个作为对照。研究了UTMD和PEI促进靶向递送和组织特异性的可行性。免疫组化检测基因沉默和凋亡诱导。电泳实验表明PEI能有效地浓缩DNA。UTMD的应用显著增加了组织转染。两种表达载体均显示基因表达存在于接受超声暴露的肿瘤的所有切片中，但在对照肿瘤中不存在。更重要的是，转基因表达的增加与PEI存在下的UTMD显著相关。沉默survivin基因可通过下调survivin和bcl-2的表达，上调bax和caspase-3的表达，有效诱导细胞凋亡。这种非侵入性的，新的UTMD与PEI的组合可以增强靶向基因递送和基因在肿瘤异种移植物中的表达，在静脉内给药，有效地没有引起任何明显的不良反应，可能是一个有希望的候选基因治疗。这种非病毒技术可以促进用shRNA沉默Survivin基因的表达，并导致显著的细胞凋亡。
Noninvasive and tissue-specific technologies of gene transfection would be valuable in clinical gene therapy. This present study was designed to determine whether it could enhance gene transfection in vivo by the combination of ultrasound-targeted microbubble destruction (UTMD) with polyethylenimine (PEI) in tumor xenografts, and illuminate the effects of gene silencing and apoptosis induction with short hairpin RNA (shRNA) interference therapy targeting human survivin by this novel technique. Two different expression vectors (pCMV-LUC and pSIREN) were incubated with PEI to prepare cationic complexes (PEI/DNA) and confirmed by the gel retardation assay. Human cervical carcinoma (Hela) tumors were planted subcutaneously in both flanks of nude mice. Tumor-bearing mice were administered by tail vein with PBS, plasmid, plasmid and SonoVue microbubble, PEI/DNA and SonoVue microbubble. One tumor was exposed to ultrasound irradiation, while the other served as control. The feasibility of targeted delivery and tissue specificity facilitated by UTMD and PEI were investigated. Moreover, immunohistochemistry analyses about gene silencing and apoptosis induction were detected. Electrophoresis experiment revealed that PEI could condense DNA efficiently. The application of UTMD significantly increases the tissue transfection. Both expression vectors showed that gene expressions were present in all sections of tumors that received ultrasound exposure but not in control tumors. More importantly, the increases in transgene expression were related to UTMD with the presence of PEI significantly. Silencing of the survivin gene could induce apoptosis effectively by downregulating survivin and bcl-2 expression, also cause up-regulation of bax and caspase-3 expression. This noninvasive, novel combination of UTMD with PEI could enhance targeted gene delivery and gene expression in tumor xenografts at intravenous administration effectively without causing any apparently adverse effect, and might be a promising candidate for gene therapy. Silencing of survivin gene expression with shRNA could be facilitated by this non-viral technique, and lead to significant cell apoptosis.
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Survivin 是预后不良的乳腺癌患者短期生存的独立预测因子。
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8.8
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